Dna hypermethylation brain cancer markers

ABSTRACT

One aspect of the present disclosure relates to a composition comprising a DNA hypermethylation brain cancer marker on acellular DNA of a human subject. DNA hypermethylation brain cancer markers (or DNA hypermethylation markers) are frequently methylated in the DNA of brain cells and acellular DNA of individuals suffering from brain cancer. These DNA hypermethylation markers are not frequently methylated in the DNA of brain cells and acellular DNA of individuals who do not suffer from brain cancer. Another aspect of the present disclosure relates to a method of diagnosing brain cancer in a human subject comprising providing a sample containing acellular DNA from the human subject, determining whether one or more DNA hypermethylation markers on the acellular DNA are hypermethylated, and diagnosing brain cancer when the DNA hypermethylation markers are hypermethylated.

PRIORITY CLAIM

The present application claims priority to U.S. Provisional PatentApplication No. 61/316,324, filed Mar. 22, 2010, which is incorporatedherein by reference.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

The invention was made with Government support under Grant No. CA084469awarded by the National Institutes of Health (NIH). The Government hascertain rights in the invention.

BACKGROUND OF THE INVENTION

Human solid tumors are characterized by a broad spectrum of genetic andepigenetic alterations. Astrocytomas are the most common type of humangliomas, and are primary malignancies in the central nervous system.These tumors are classified by varying degrees of malignancy with gradeIV astrocytomas (glioblastomas) being the most aggressive. Animal modelsand recent progress in stem cell research have now established thatdividing cells in the subventricular zone (SVZ) give rise toastrocytomas (1). Human cancer genome sequencing studies have cataloguedcharacteristic mutations in glioblastomas. These studies have uncoveredthat individual tumors generally have ˜30-40 mutations in gene codingsequences that alter the amino acid composition of a protein (2). Themost commonly mutated or homozygously deleted genes include CDKN2, TP53,EGFR, PTEN, NF1, PIK3CA, and IDH1. At the epigenetic level, tumorspecimens are most often characterized in terms of altered DNAmethylation (3). Aberrations in DNA methylation patterns may havecritical effects on tumor initiation and progression (4). Both DNAhypermethylation of specific genes and DNA hypomethylation commonlyaffecting repetitive DNA are observed in brain cancers (5-11). CpGisland hypermethylation can affect a variety of different genes, rangingfrom regulators of cell cycle progression and apoptosis to DNA repairgenes and developmental transcription factors. With regard to the latterclass of genes, it has recently been recognized that a large fraction ofthe genes commonly methylated in human tumors represent targets of thePolycomb complex in human embryonic stem cells (12-17). In the presentdisclosure, we have employed a comprehensive methylation profilingtechnique, developed in our laboratory, and termed “methylated-CpGisland recovery assay” or MIRA (12). This assay was used in conjunctionwith CpG island and promoter microarrays (MIRA-chip) to characterize theCpG island methylome in normal human brain tissue and brain tumors(astrocytomas) representing different stages of tumor progression andmalignancy.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS AND TABLES

FIG. 1: Localization of methylated CpG islands in normal human brainrelative to known genes and correlation between gene methylation andgene expression levels.

A. Localization of the methylation peaks in normal brain relative togene position. Each normal brain sample had between 6,000 and 7,000methylation peaks. B. Correlation between 5′ gene end methylation innormal brain and gene expression levels. Public microarray geneexpression data of whole brain tissue were obtained from the GNFSymAltas database. Genes with methylation peaks at their 5′ end aregenerally expressed at lower level than genes without methylation peaksat the 5′ end (Kolmogorov-Smirnov Test P value=1.8e-10). C. Correlationbetween gene body methylation in normal brain and gene expressionlevels. Genes with intragenic methylation peaks are generally expressedat a higher level than genes without intragenic methylation(Kolmogorov-Smirnov Test P value <2.2e-16).

FIG. 2: Confirmation of tumor-associated DNA methylation by combinedbisulfite restriction (COBRA) analysis and sodium bisulfite sequencing.The CpG island associated with the BCL2L11 gene was analyzed in normalbrain DNA (N) and in brain tumors (numbered). A. COBRA assays. In theCOBRA assay, cleavage by BstUI indicates methylation at 5′CGCGsequences. B. Bisulfite sequencing. In the bisulfite sequencing panels,open circles represent unmethylated CpG sites and closed circlesrepresent methylated CpG sites. The triangles indicate BstUI sites.

FIG. 3: Tumor-specifically methylated genes and tumor grade. A. Thenumber of methylated regions is plotted according to tumor grade.Tumor-specific methylation peaks were identified. These peaks are notfound in any of the six normal samples. The grey horizontal linesrepresent the median number in each grade. B. Cluster analysis.Methylation peaks commonly found in tumor samples but not in normalsamples were identified (see detailed description). Then the data weresubjected to hierarchical clustering with Euclidean distance and averagelinkage method using Cluster v3.0 and visualized in Java tree-view.Light grey color represents a methylated state, and black represents anunmethylated state.

FIG. 4: Clusters of hypermethylated CpG islands at the HOXD locus. Thetop of the Figure indicates the location of the individual HOXD genes.The associated CpG islands are shown as dark boxes. The bottom part ofthe Figure shows the NimbleGen array data (methylated fraction versusinput) for three normal brain tissues (N) and three grade IIIastrocytomas (T). The methylation signal is shown plotted along thechromosome as a P value score. Therefore, the minimum number on they-axis is 0 (when P=1). The P value score was obtained by NimbleScansoftware and is derived from the Kolmogorov-Smirnov test comparing thelog 2 ratios (MIRA versus input) within a 750 bp window centered at eachprobe and the rest of the data on the array.

FIG. 5: Clusters of hypermethylated CpG islands at the histone cluster 1and HLA loci. The top of the Figure indicates the location of thehistone cluster 1 and MHC genes. The location of Refseq genes and mRNAsis according to the UCSC Genome Browser. The associated CpG islands areshown as vertical bars. The bottom part of the Figure shows theNimbleGen array data (methylated fraction versus input) for three normalbrain tissues (N) and three grade III astrocytomas (T). At this level ofresolution, each bar represents a methylated CpG island. The methylationsignal is shown plotted along the chromosome as a P value score.

FIG. 6: Confirmation of tumor-associated DNA methylation by combinedbisulfite restriction (COBRA) analysis and sodium bisulfite sequencing.The CpG island associated with the ZNF274 gene was analyzed in normalbrain DNA (N) and in brain tumors (numbered). A. COBRA assays. In theCOBRA assay, cleavage by BstUI indicates methylation at 5′CGCGsequences. B. Bisulfite sequencing. In the bisulfite sequencing panels,open circles represent unmethylated CpG sites and closed circlesrepresent methylated CpG sites. The triangles indicate BstUI sites.

FIG. 7: Confirmation of tumor-associated DNA methylation by combinedbisulfite restriction (COBRA) analysis and sodium bisulfite sequencing.The CpG island associated with the RAX gene was analyzed in normal brainDNA (N) and in brain tumors (numbered). A. COBRA assays. In the COBRAassay, cleavage by BstUI indicates methylation at 5′CGCG sequences. B.Bisulfite sequencing. In the bisulfite sequencing panels, open circlesrepresent unmethylated CpG sites and closed circles represent methylatedCpG sites. The triangles indicate BstUI sites.

FIG. 8: Overlap of the most commonly methylated genes and gene markingby the Polycomb complex. Methylation data is for the 288 commonlymethylated genes, i.e. those methylated in >25% of the tumors. Data forPolycomb marking as indicated by occupancy with H3K27me3 in embryonicstem cells was obtained from Lee et al., Cell 2006; 125:301-313. Theoverlap is statistically highly significant (P<2.2e-16).

FIG. 9: Clusters of hypermethylated CpG islands at the HOXC locus. Aview of the UCSC Genome Browser is shown at the top indicating thelocation of the individual HOXC genes. The associated CpG islands areshown as vertical bars. The bottom part of the Figure shows theNimbleGen array data (methylated fraction versus input) for three normalbrain tissues (N) and three grade III astrocytomas (T). The methylationsignal is shown plotted along the chromosome as a P value score.

FIG. 10: Clusters of hypermethylated CpG islands at the BARHL2 locus. Aview of the UCSC Genome Browser is shown at the top indicating thelocation of the gene. The associated CpG islands are shown as boxes. Thebottom part of the Figure shows the NimbleGen array data (methylatedfraction versus input) for three normal brain tissues (N), three gradeIII astrocytomas (top three T scans) and three glioblastomas (bottomthree T scans). The methylation signal is shown plotted along thechromosome as a P value score.

FIG. 11: Clusters of hypermethylated CpG islands at the PITX2 locus. Aview of the UCSC Genome Browser is shown at the top indicating thelocation of the gene. The associated CpG islands are shown as boxes. Thebottom part of the Figure shows the NimbleGen array data (methylatedfraction versus input) for three normal brain tissues (N), three gradeIII astrocytomas (top three T scans) and two glioblastomas (bottom two Tscans). The methylation signal is shown plotted along the chromosome asa P value score.

FIG. 12: Clusters of hypermethylated CpG islands at the SIM1 locus. Aview of the UCSC Genome Browser is shown at the top indicating thelocation of the gene. The associated CpG islands are shown as boxes. Thebottom part of the Figure shows the NimbleGen array data (methylatedfraction versus input) for three normal brain tissues (N) and threegrade III astrocytomas (T). The methylation signal is shown plottedalong the chromosome as a P value score.

FIG. 13: Clusters of hypermethylated CpG islands on chromosome 14including several homeobox genes. A diagram at the top indicates thelocation of the genes with chromosomal coordinates. A close-up view isshown in the lower part of the Figure. The bottom parts of the Figurepanels show the NimbleGen array data (methylated fraction versus input)for three normal brain tissues (N) and three grade III astrocytomas (T).The methylation signal is shown plotted along the chromosome as a valuescore.

FIG. 14: Clusters of hypermethylated CpG islands on chromosome 1spanning an area of segmental duplications. A view of the UCSC GenomeBrowser is shown at the top indicating the location of the individualgenes. CpG islands are shown as vertical bars. Segmental duplicationswith >90% similarity to other chromosomal segments are shown as stripedgrey or black bars. Information on gene order and duplications was takenfrom the UCSC Genome Browser, March 2006 assembly(http://genome.ucsc.edu/cgi-bin/hgGateway). The bottom part of theFigure shows the NimbleGen array data (methylated fraction versus input)for three normal brain tissues (N) three grade III astrocytomas (topthree T scans) and three glioblastomas (bottom three T scans). At thislevel of resolution, each bar represents a methylated CpG island. Themethylation signal is shown plotted along the chromosome as a P valuescore.

FIG. 15: Model for astrocytoma initiation in neural stem cells. Thecontribution from DNA hypermethylation events is the permanent silencingof neuronal differentiation genes thus locking the stem cells and theirderivative transitamplifying cells into a prolonged undifferentiatedstate and allowing additional oncogenic events to occur (either geneticor epigenetic) leading to initiation of malignant transformation andprogression towards astrocytoma.

Table 1: Frequently methylated regions in brain

Table 2: Frequently methylated regions in grade I tumors.

Table 3: Functional annotation of 5′-end methylated genes in tumorsamples.

Table 4: Genes methylated at the 5′ end in normal brain.

Table 5: Frequently methylated 5′ gene ends in brain tumors.

Table 6: Genes with frequent hypomethylation at the 5′ end.

DETAILED DESCRIPTION OF THE INVENTION

One aspect of the present disclosure relates to a composition of mattercomprising a DNA hypermethylation brain cancer marker on acellular DNAof a human subject. DNA hypermethylation brain cancer markers (or DNAhypermethylation markers) are frequently methylated in brain cancer andnot frequently methylated in normal brain. Some examples of DNAhypermethylation markers are listed in Table 1. In certain embodiments,DNA hypermethylation markers include, without limitation, DNA sequenceshaving SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ IDNO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8, SEQ ID NO. 9, SEQ IDNO. 10 and combinations thereof. In another embodiment, the DNAhypermethylation markers which are frequently methylated in grade Ibrain tumors, including EBF1, include DNA sequences having SEQ ID NO. 2,SEQ ID NO. 11, SEQ ID NO. 12, SEQ ID NO. 13, SEQ ID NO. 14, SEQ ID NO.15, SEQ ID NO. 16, SEQ ID NO. 17, and combinations thereof (Table 2).

In certain embodiments, a DNA hypermethylation marker comprises adjacentCpG islands (“CpG clusters”) that are frequently methylated in braincancer. In certain embodiment, the DNA hypermethylation marker comprisesfive or more adjacent CpG islands that underwent frequent simultaneoushypermethylation in brain cancer but are not frequently methylated innormal brains. In certain embodiments, the brain cancer is a brain tumor(e.g. a grade I, grade II, grade III or grade IV tumor). In certainembodiments, the DNA hypermethylation marker comprises CpG clustersassociated with homeobox genes. In certain embodiments, the DNAhypermethylation marker comprises CpG clusters that are not associatedwith homeobox genes. Examples of CpG clusters associated with homeoboxgenes include, without limitation, the HOX gene clusters HOXD (FIG. 4),HOXC (FIG. 9), HOXA, HOXB, the homeobox genes DLX1, BARHL2 and PITX2(FIGS. 10 and 11), developmental transcription factor gene simple-mindedone (SIM1) (FIG. 12), CpG clusters on chromosome 14 (FIG. 13) whichcomprises several homeobox and transcription factor genes includingNKX2-8 PAX9 and FOXA1. Another example of CpG clusters is on chromosome6p21 (25,700,000-30,500,000) (FIG. 5), comprising the major histonecluster 1 and several HLA genes of the MHC locus (FIG. 5).

Another aspect of the present disclosure relates to a method ofdiagnosing brain cancer in a human subject comprising:

providing a sample containing acellular DNA from the human subject,wherein:

-   -   the sample is selected from the group consisting of a blood        sample, serum sample and plasma sample; and        determining whether one or more DNA hypermethylation markers on        the acellular DNA are hypermethylated, wherein:    -   the hypermethylation of the DNA hypermethylation markers is        indicative of brain cancer, and    -   the DNA hypermethylation markers are selected from the group        consisting of SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID        NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8,        SEQ ID NO. 9, SEQ ID NO. 10, and combinations thereof.

In certain embodiments, the brain cancer diagnosed is a brain tumor. Incertain embodiments, the brain tumor is a grade I to grade IV tumor. Incertain embodiments, the brain tumor is a grade I tumor. In certainembodiments, the DNA hypermethylation markers are selected from thegroup consisting of SEQ ID NO. 2, SEQ ID NO. 11, SEQ ID NO. 12, SEQ IDNO. 13, SEQ ID NO. 14, SEQ ID NO. 15, SEQ ID NO. 16, SEQ ID NO. 17, andcombinations thereof.

The present disclosure provides a global view of DNA hypermethylationevents occurring in human astrocytomas. We determined that 6,000 to7,000 CpG islands are methylated in normal brain. This number representswell over 20% of all CpG islands in total and ˜5% of all CpG islands at5′ gene ends were methylated. We found that genes with methylation peaksat their 5′ end are expressed at significantly lower level than geneswithout methylation peaks at the 5′ end.

One important attribute of a diagnostic for brain cancer is that the DNAhypermethylation markers are that they should be methylated in nearlyall tumors, including early stage and low-grade tumors. To acceleratebiomarker development, we created a list of 428 methylated regions thatwere methylated in >25% of tumors (i.e. in at least 8 of the 30 tumorsanalyzed) (Table 1). The most frequently methylated region was anuncharacterized CpG island on chromosome 7 that is methylated in 87% ofthe tumors (SEQ ID NO. 1: Chromosome 7, starting from 152252636 to152253475). The most frequently methylated gene-associated region waswithin the gene body of the EBF1 gene located on chromosome 5 (SEQ IDNO. 2: Chromosome 5, starting from 158456493 to 158457132). The secondmost frequently methylated gene was DHH (Desert hedgehog), methylated in83% of the tumors (SEQ ID NO. 3: Chromosome 12, starting from 47771091to 47771619). We identified methylated regions that weremethylation-positive in at least 5 of the 6 grade I tumors, includingEBF1 (Table 2).

One somewhat unexpected result of our study was the finding that gradeIV tumors have fewer hypermethylated regions than several of the gradeII and III tumors.

Several known tumor suppressor genes are methylated in astrocytomas.These include the genes CDKN2A, PTEN, p14ARF, and RB. The reason thatthese genes are not listed in Tables 1 and 5 are for the most part thatthese genes were not methylated in more than 25% of the tumors (ourcriterion to be included on the list). This finding is consistent withthe relatively infrequent methylation of these genes reported in theliterature. We found methylation of the PTEN promoter in 20% of thetumors. CDKN2A was methylated in 23% and two tumors displayed no signalin the entire gene area suggesting homozygous deletion. For the MGMTgene, for which reported methylation frequencies in gliomas generallyrange from 40% to over 70%, we found weak signals at the promoter innormal brain and enhanced signals in about 40% of the tumors. However,the statistical cutoff used to score differential peaks (see Methods)did not allow this gene to be scored on our list. On the list of the 288most frequently methylated genes, homeobox genes and other developmentalregulatory transcription factors were greatly overrepresented.Thirty-one percent of these genes were targets of the Polycomb complex(H3K27me3-marked) in embryonic stem cells (FIG. 8).

One important finding of our studies was the frequent occurrence ofclusters of adjacent hypermethylated CpG islands. Several examples ofthese included homeobox genes. Mechanistically, this phenomenon islikely related to occupancy of larger DNA sequence stretches by thePolycomb complex surrounding homeobox genes, and would, again, reflectPolycomb-targeted methylation but in a more large-scale fashion. We alsoidentified another situation in which multiple adjacent CpG islandsbecame hypermethylated in tumors, as a result of being present as partof multi-copy gene families. The two examples observed, reflectingextensive hypermethylation of the NBPF gene family and of CpG islands inhistone cluster 1 and HLA genes in the MHC cluster suggest that it isgene duplication events per se that may trigger DNA hypermethylation.Paradoxically, this situation is the opposite of what we have observedin lung squamous cell carcinomas in which segmental duplications aregenerally hypomethylated in tumors (22). These data suggest thatdifferent types of malignancies have different aberrations concerningmethylation of duplicated gene sequences.

Gene ontology analysis indicated that a significantly enriched genecategory with common 5′ gene end methylation included genes involved inbrain development and neurogenesis, once again with frequent involvementof Polycomb target and homeobox genes (Table 3). Recent mouse modelshave shown that gliomas arise from neural stem cells in thesubventricular zone (SVZ) and/or transit-amplifying progenitors ratherthan from normal glial progenitors or oligodendrocytes (26, 27). Neuralstem cells in the adult SVZ give rise to highly proliferatingtransit-amplifying progenitor cells, which then differentiate into thedifferent lineages, including neuroblast, astrocyte and oligodendrocyteprecursors. Aberrant or failed neurogenesis, due to methylation-inducedpermanent silencing of neuronal differentiation genes such as POU4F3,GDNF, OTX2, NEFM, CNTN4, OTP, SIM1, FYN, EN1, CHAT, GSX2, NKX6-1, PAX6,RAX, DLX2, and BMP4 and several others (Table 3) may be a fundamentalcomponent of glioma initiation from neural stem cells (see model in FIG.15). For example, BMP4 has been shown to reduce the glioma-initiatingcell pool in glioblastomas (28). Given that SVZ neural stem cells arethe cells of origin of gliomas (26, 27), these methylation changes donot simply reflect lineage committed oligodendrocyte progenitors asglioma precursors, in which such genes are likely methylation-silenced.Rather, aberrant methylation of a group of genes involved in neuronaldifferentiation, in cooperation with other oncogenic events such as TP53mutation, may shift the balance from regulated differentiation towardscell proliferation and gliomagenesis in neural stem cells. The permanentsilencing of pro-differentiation genes may lead to the accumulation of acell population unable to differentiate that can persist long enough,perhaps over the lifetime of an individual, to acquire the necessarytransforming genetic or epigenetic aberrations. Our data suggest thatepigenetic differentiation therapy might provide a useful approach forthe management of this lethal disease.

Tissue and DNA Samples.

Six normal human brain tissues from accident victims were obtained fromCapital Biosciences (Gaithersburg, Md.) and BioChain (Hayward, Calif.).Twenty-four brain tumor tissues from astrocytomas (World HealthOrganization grade I-IV) were obtained on Institutional Review Boardapproved protocols from randomly selected patients who had brain tumorsurgery at the Department of Neurosurgery at the University Hospital inDresden during 1997-2007. Histological diagnosis was performed by alocal neuropathologist and confirmed by reference neuropathology. Thesetumors are labeled T1-(1-6), T2-(1-6), T3-(1-6), and T4-(1-6), where thenumber immediately following the T reflects tumor grade. Six tumorslabeled T2-7, T3-(7-9), and T4-(7-8) were astrocytomas obtained fromAsterand (Detroit, Mich.). No patient had chemotherapy before tissuesampling.

Methylated-CpG Island Recovery Assay (MIRA) and MicroarrayHybridization.

Tumor and normal brain DNA was fragmented by sonication to ˜500 bpaverage size on agarose gels. Enrichment of the methylateddouble-stranded DNA fraction by MIRA was performed as describedpreviously (18, 19). The labeling of amplicons, microarrayhybridization, and scanning were performed according to the NimbleGenprotocol. NimbleGen tiling arrays were used for hybridization (385KHuman CpG Island plus Promoter arrays). The single array design coversall 28,226 UCSC Genome Browser-annotated CpG islands and the promoterregions for all RefSeq genes. The promoter region covered is onekilobase (−800 to +200 relative to the transcription start sites). Forall samples, the MIRA enriched DNA was compared with the input DNA.

Identification and Annotation of Methylated Regions.

Log 2 ratio data were converted into P value scores using theKolmogorov-Smirnov (KS) test with a 750 bp window by using theNimbleScan software (Sp=−log 10(p)). Probes were selected as positive iftheir P value scores were above 2 (P<0.01). For our analysis, we defineda methylated region of interest (methylation peak) as a region with atleast 4 consecutive positive probes (i.e. no gaps) covering a minimumlength of 350 bp. This stringent definition will give few false-positiveresults. Identified methylation peaks were mapped relative to knowntranscripts defined in the UCSC Genome Browser HG18 RefSeq database.Methylation peaks falling into 1000 bp relative to transcription startsites were defined as 5′-end peaks; methylation peaks falling within1000 bp of RefSeq transcript end sites were defined as 3′-end peaks, andthose falling within gene bodies (from 1000 bp downstream oftranscription start to 1000 bp upstream of transcript end) were definedas “intragenic” peaks. Methylation peaks that are not close to any knowntranscripts were defined as “intergenic.”

Identification of Tumor-Specific Methylated Regions

Methylation peaks in tumor samples were identified as above, andmethylation peaks in normal samples were identified using a P valuescore >=1.7 to exclude regions with weak methylation. The methylatedregions in tumor and normal samples were matched by outer join and theirmethylation status was determined by the presence of methylation peaksin each sample in these matched regions. Only the regions methylated inmore than 25% of the 30 tumors and unmethylated in all of the six normalsamples were considered as candidates. The average P value scores ofprobes within these candidates in tumor samples were compared to theaverage P value scores of probes in normal samples, and the finaltumor-specific methylation peaks were selected if the difference wasmore than 1. The tumor specific methylation peaks were also mapped tothe 28,226 CpG islands defined as such at the UCSC Genome Browser. TheCpG islands are defined as having a G/C content of >50%, CpG observed toexpected ratio of >0.6 and a length of at least 200 bp. These CpGislands were also mapped relative to known transcripts in the HG18RefSeq database using the same approach described above. The datareported in this paper have been deposited in the National Center forBiotechnology Information Gene Expression Omnibus (GEO) database(accession no. GSE19391).

DNA Methylation Analysis Using Sodium Bisulfite-Based Methods.

DNA was treated and purified with the EpiTect Bisulfite kit (Qiagen;Valencia, Calif.). PCR primer sequences for amplification of specificgene targets in bisulfitetreated DNA are available upon request. The PCRproducts were analyzed by combined bisulfite restriction analysis(COBRA) as described previously (20). Additionally, PCR products frombisulfite-converted DNA were cloned into the pDrive PCR cloning vector(Qiagen), and 8-12 individual clones were sequenced.

Analysis of DNA Methylation in Normal Brain.

MIRA-chip has proven to be a sensitive, robust and reproducibletechnique for mapping DNA methylation patterns in mammalian genomesincluding cancer tissues (12, 14, 19). Six normal brain tissue DNAsamples from accident victims were used to determine patterns ofmethylation at CpG islands and promoters using NimbleGen tiling arrays.After enrichment of the methylated DNA fractions by MIRA, MIRA and inputfractions were mixed and hybridized to the arrays, and data werenormalized and analyzed as described in Materials and Methods. We founda surprisingly large number of methylated CpG islands in normal humanbrain ranging from 6,000 to 7,000 in each sample. These numberscorrespond to approximately 21-25% of all CpG islands as defined above.We then analyzed the localization of these methylation peaks relative toknown genes. We found that 9.3% of the methylation peaks were at the 5′ends of genes (defined as +/−1000 bp relative to transcription startsites), 53.8% were within gene bodies (defined as within 1000 bpdownstream of the transcription start site and 1000 bp upstream of thegene end of RefSeq genes), and 9.1% were near the 3′ end of genes(defined as +/−1000 bp of the gene end). 27.8% of the methylation peakswere not associated with genes (FIG. 1A).

Next, we focused on those genes with methylation peaks at the 5′ end andexplored the relationship of this 5′ gene end methylation and geneexpression patterns using public microarray gene expression data ofwhole brain tissue from the GNF SymAltas database. There were 456 geneswith 5′-end methylation in all normal samples (Table 4). We found thatgenes with methylation peaks at their 5′ end are generally expressed atlower levels than genes without methylation peaks at the 5′ end, and thedifference is highly significant (Kolmogorov-Smirnov Test Pvalue=1.8e-10) (FIG. 1B). Using the Database for Annotation,Visualization and Integrated Discovery (DAVID, version 6), we obtainedannotation information for 377 of these genes. Functional annotationanalysis showed that genes involved in spermatogenesis are highlyenriched (Benjamini-corrected P value=7.28e-04). No other gene ontologycategories had corrected P values <0.05.

We then looked at the genes with methylation peaks within the gene body.There were 1768 genes with gene body methylation and without 5′-endmethylation. Consistent with previous findings in B cells (21, 19), weobserved that genes with gene body methylation are generally expressedat higher levels than genes without gene body methylation (FIG. 1C).

Analysis of DNA Methylation in Brain Tumors Shows Frequent Methylationof Genes Involved in Neuronal Differentiation.

In order to characterize methylation patterns in human brain tumors, weperformed MIRA-chip analysis on 30 brain tumors including six grade I,seven grade II, nine grade III, and eight grade IV astrocytomas(glioblastomas). To confirm that the microarray data correctly reflecttrue methylation differences, we analyzed several genes includingBCL2L11, RAX, FOXP4, and ZNF274 bp sodium bisulfite-based DNAmethylation analysis techniques. Consistent with previous datademonstrating the reliability of the MIRA assay (12, 14, 22, 19), weconfirmed methylation differences between normal and tumor tissues byCOBRA analysis and by bisulfite sequencing, as shown for thepro-apoptotic gene BCL2L11 also known as BIM (FIG. 2), for the RAX andZNF274 genes (FIGS. 6 and 7) and for FOXP4. The BCL2L11 gene wasmethylated in 21 of the 30 brain tumors and mostly in tumors of grade IIand higher (FIG. 2).

We catalogued those genes that are methylated in the brain tumors anddefined a tumor-specifically methylated gene as one that was notmethylated in any of the six normal brain tissue DNAs. Using thesecriteria, we determined the number of tumor-methylated regions in eachtumor as shown in FIG. 3A. The number of tumor-specifically methylatedregions increased in astrocytomas going from grade Ito grade III.However, this number, on average, decreased in grade IV tumors relativeto grade II and III tumors. Four of the normal brain tissues availableto us were from the frontal lobe, one was from the occipital lobe andone from an unspecified region of the brain. Among these six normalbrain tissues, there were very few differences in terms of number andlocation of methylated CpG islands. Astrocytomas came from severaldifferent regions of the brain including frontal, parietal, occipital,fronto-temporal, and temporal lobes, and three of the stage I tumorswere from the cerebellum. When looking at location vs. number ofmethylation targets adjusting for tumor stage, there was no significantcorrelation, but the numbers of tumors are too small to exclude thepossibility of brain region-specific differences in tumor-associatedmethylation. We then established a list of genes that are most commonlymethylated across different tumor specimens. These genes areunmethylated, i.e. lack a peak on the arrays in all six normal brainsamples, and are methylated in at least 25% of the tumors (i.e. in atleast 8 out of 30 tumors). We compiled a list of 428 methylation peaksthat fit these criteria (Table 1). A hierarchical clustering of thesetumor-specific methylated regions is shown in FIG. 3B. Grade I tumorsform a tight cluster, adjacent to 5 higher grade tumors, T4-2, T3-3,T2-4, T4-5, and T4-1. The other higher-grade tumors form two distinctclusters. One cluster includes T2-2, T2-3, T3-2, T2-5, T3-4 and T2-6.The rest of the tumors form another cluster and they have the mostmethylation peaks. Of the 428 frequently methylated regions, 403 overlapwith CpG islands, and 288 fall into genes, i.e. they are within 1000 bpupstream of the transcription start site and 1000 bp downstream of thetranscription end of a gene. There were 174 methylation peaks that canbe annotated to 5′-ends of Refseq transcripts (Table 5). Of these, 164genes had annotation information in DAVID. Functional analysis revealedthat genes involved in organ, nervous system and brain development, andneurogenesis are highly enriched (Table 3). Many of these genes aretranscription factor and homeobox genes. An analysis of Interpro domainsrevealed 35 homeobox genes in the list of 288 genes with methylationpeaks (Enrichment-fold=12.2, P=3.7e-26, InterPro domain IPR001356).Since homeobox genes and other developmental regulatory genes are knowntargets of the repressive Polycomb complex in embryonic stem (ES) cells,we compared the list of commonly methylated genes with the list of genesmarked as Polycomb targets in human ES cells (23). FIG. 8 shows thatindeed 30.7% of the methylated genes are Polycomb targets. The observednumber of overlapping targets between MIRA and Polycomb is more than7-fold higher than expected by chance, indicating a strong enrichment(P<2.2e-16; Fisher's exact test).

Although we focused mostly on DNA hypermethylation in this study, wealso looked for hypomethylation of CpG islands in the tumor samplesrelative to normal brain. We identified a list of 46 genes where thehypomethylation event is frequently present at the 5′ gene end (Table6). This list was enriched for cancer-testis antigen genes including thegenes SOHLH2, SSX2, SSX4B, SSX8, SSX9, and PAGES. This class of genes isnormally expressed in germ cells, is silenced by DNA methylation insomatic tissues but can be reactivated and demethylated in cancer.

Clusters of CpG Island Hypermethylation in Brain Tumors.

In addition to the many singular hypermethylated CpG islands presentalong all chromosomes, we identified a number of prominent clusters ofhypermethylated CpG islands. These clusters were defined as a minimum offive adjacent CpG islands that underwent simultaneous hypermethylationin at least five of the tumors but are methylation-free in all normalbrains. These clusters could be divided into two categories: (1) CpGislands associated with homeobox genes and (2) hypermethylation clustersnot associated with homeobox genes. Category 1 included the followingloci: the HOX gene clusters HOXD (FIG. 4) and HOXC (FIG. 9), and thehomeobox genes DLX1, BARHL2 and PITX2 (FIGS. 10 and 11), as well as thedevelopmental transcription factor gene simple-minded one (SIM1) (FIG.12). The HOX clusters HOXA and HOXB were also hypermethylated in tumorsbut several of the genes in these two clusters had considerablemethylation in normal brain. The hypermethylated CpG islands at thesehomeobox gene loci were found not only at the 5′ gene ends but alsowithin the gene bodies and at the 3′ ends. Chromosome 14 shows aparticularly illustrative example of homeobox gene-associatedhypermethylation of CpG island clusters (FIG. 13). In this case, severalhomeobox and transcription factor genes, including NKX2-8, PAX9, andFOXA1 are located within a 1.8 Mb segment, and no less than 24contiguous CpG islands become hypermethylated in astrocytomas. A 360kilobase stretch at chromosome 1q21.2 (145,940,000-146,300,000)contained 12 hypermethylated CpG islands. In this case, four consecutivetumor-specifically hypermethylated islands and eight adjacenttumorspecifically methylated islands were separated by two CpG islandsthat were methylated in tumors and in normal brain. Interestingly, thischromosomal region is rich in segmental duplications (FIG. 14). Thislocus contains the neuroblastoma breakpoint family (NBPF) of genes namedafter a chromosomal translocation found in a neuroblastoma patient (24).The NBPF genes encode proteins of unknown function and have a repetitivestructure with high intragenic and intergenic sequence similarity inboth coding and noncoding regions. No discernable NBPF orthologues havebeen identified in the genomes of mouse or rat. This gene family showsprimate-specific duplications that result in species-specific arrays ofNBPF homologous sequences (24). The hypermethylation cluster onchromosome 6p21 (25,700,000-30,500,000) spanning 4.8 megabases isparticularly striking. This hypermethylation cluster was present in 16out of 30 tumors analyzed (FIG. 5). The region contains the majorhistone cluster 1 and several HLA genes of the MHC locus. This largecluster of histone genes contains 55 histone genes (25). Thus, thischromosomal region represents a segment where closely related genes, thehistone gene families and HLA genes are present and are subject tohypermethylation in brain tumors. Although some CpG islands in thisregion are methylated in normal brain as well, there are 42 CpG islandsthat are subject to tumor-specific hypermethylation.

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TABLE 1 Frequently methylated regions in brain tumors. Peaks Average PNo. of Tumors Downstream Gene Within chr Start End value score * withPeaks Strand Start End Symbol Strand Start chr1 2456587 2457246 7.35 19chr1 37973297 37974046 12.12 17 − 37954232 chr1 38002438 38003587 10.1012 chr1 46712589 46713323 7.31 9 chr1 47469314 47470340 8.43 11 chr154594420 54595152 6.92 14 − 54464782 chr1 60311689 60312524 9.01 16 chr163556597 63560345 8.49 15 chr1 90963290 90965440 10.04 16 chr1 111614814111615476 11.27 20 chr1 119328183 119328832 10.62 21 − 119227188 chr1143012194 143012944 10.97 14 − 143011875 143053112 LOC728875 − 143011875chr1 144674586 144675239 11.02 17 chr1 144690585 144691329 9.13 17 chr1144702031 144702576 7.97 15 chr1 145971587 145972240 11.05 19 chr1145987585 145988329 9.59 19 chr1 145999042 145999587 8.90 16 chr1146184793 146185549 8.70 19 chr1 146219303 146219957 9.13 12 chr1146248547 146249292 7.11 12 chr1 146256160 146256719 8.21 16 chr1146466570 146467421 9.58 15 chr1 147599481 147600124 13.18 21 chr1147600226 147600785 12.95 17 chr1 147665315 147665969 7.78 12 chr1147882409 147883136 6.55 15 + 147843076 147883408 LOC728855 chr1147985011 147986318 13.04 21 chr1 150078663 150079332 11.19 11 +150077568 chr1 150349668 150350912 10.81 12 − 150345416 chr1 153320294153321238 8.12 11 + 153317971 chr1 153557083 153557827 8.77 11 −153556874 153560562 C1orf104 chr1 154527610 154528254 8.01 14 −154529107 154532073 C1orf85 + 154520693 chr1 155085184 155085851 7.40 16− 155077288 chr1 165119820 165120544 12.95 12 chr1 179719350 17971990410.69 13 chr1 181421423 181422187 9.18 19 chr1 196156945 196157494 11.0720 + 196153139 chr1 207914587 207915953 6.93 9 + 207915292 207916358G0S2 + 207915292 chr1 219131519 219132268 10.21 20 chr1 219134095219134834 6.51 11 chr1 222002983 222003622 6.09 15 + 221966741 chr1222870205 222872650 9.25 10 + 222870801 chr1 226700008 226700767 11.8610 chr1 234752777 234754328 9.67 11 + 234748187 chr10 7489395 74901539.35 15 − 7244254 chr10 46412477 46413703 7.66 21 chr10 5048654650487915 9.95 11 chr10 50488946 50491715 10.13 15 + 50488352 50490772SLC18A3 + 50488352 chr10 79066119 79066773 9.27 12 − 78314641 chr1081732969 81733640 6.61 9 chr10 99070134 99071063 10.07 15 + 9906901199071662 FRAT1 + 99069011 chr10 102580005 102580574 5.67 12 + 102495457102579688 PAX2 chr10 102972304 102973663 11.25 18 chr10 120344105120345734 7.81 11 − 120342905 chr10 124628733 124629607 7.80 12 chr111718992 1720660 11.14 21 − 1447268 chr11 1721203 1723357 8.45 9 chr111727398 1728562 7.56 12 chr11 2102906 2103345 7.82 16 chr11 21831942184458 6.81 9 chr11 10428000 10428654 7.76 16 chr11 14883948 148843028.94 20 chr11 19220281 19221020 7.43 18 chr11 22319354 22320095 12.0013 + 22316242 chr11 31789500 31795000 8.20 10 − 31768059 chr11 3241145332412722 13.77 15 − 32365900 chr11 44283814 44284478 7.06 13 − 44242733chr11 44935052 44935706 8.16 17 chr11 63531706 63532455 8.98 14 −63522606 chr11 63823533 63825290 17.92 13 + 63815368 63824079 KCNK4chr11 64947186 64947820 7.85 12 chr11 73049464 73050318 8.48 16 +73037382 chr11 94523361 94524500 13.90 19 chr11 124133793 124135432 6.7613 − 124128228 chr11 133762389 133763138 6.79 15 − 133753607 chr126803351 6804025 5.33 12 + 6801223 chr12 21985422 21986671 6.67 11 chr1242231009 42232738 12.35 17 chr12 45511201 45511951 7.23 13 chr1247771091 47771619 9.92 24 − 47769472 chr12 47975997 47977646 11.74 21 +47975175 chr12 52418168 52418832 5.91 8 chr12 52646454 52647108 9.76 19− 52642362 chr12 53049097 53049742 7.62 18 − 53049186 53071350 ZNF385Achr12 55168271 55169242 10.27 19 chr12 55918204 55919848 21.03 9 −55914952 chr12 56300983 56301939 13.42 12 + 56299959 chr12 7962616979626805 12.87 20 + 79625576 79627382 MYF6 + 79625576 chr12 8419722984198765 8.80 11 chr12 101868310 101868969 7.61 12 chr12 113322635113323304 9.71 14 − 113288284 chr12 115800795 115801644 9.95 17 −115783409 chr13 27399628 27401487 14.44 11 chr13 40928925 40929564 6.5017 chr13 52211145 52212607 7.69 14 chr13 102224101 102224965 10.63 16chr13 112725390 112726034 8.32 17 + 112681655 chr14 22360909 223613658.50 24 chr14 23115638 23116287 7.60 14 − 23107083 chr14 3606137036062206 8.09 12 chr14 36122209 36123483 8.76 9 chr14 36194850 3619549912.01 15 chr14 36196539 36197988 8.90 13 + 36196532 chr14 3620153936202583 10.95 15 + 36196532 chr14 37137508 37138866 5.61 11 chr1437748210 37750570 10.41 18 + 37746954 chr14 53488199 53488653 10.84 20 −53486204 chr14 53491636 53492707 12.27 14 − 53486204 chr14 5349983553500364 16.29 12 chr14 56344682 56346630 8.36 9 − 56337177 chr1460021741 60022400 6.81 12 chr14 60043368 60044022 9.32 12 chr14 6004509960047942 9.30 11 + 60045690 60048278 SIX6 + 60045690 chr14 7377645073777601 6.38 8 + 73775927 chr14 74147083 74147922 9.41 19 − 74034638chr14 94304390 94305319 7.99 17 − 94304312 94306252 GSC − 94304312 chr1494308930 94310289 11.73 18 chr14 105023673 105024599 8.21 12 chr1543195869 43196818 10.16 13 − 43196977 43209349 DUOXA1 + 43193815 chr1543783619 43784273 7.10 18 chr15 50867775 50868924 6.92 18 − 50836644chr15 50873886 50875041 9.91 16 chr15 62984637 62985202 10.84 19 chr1565913980 65915431 8.60 12 + 65904994 65913228 LBXCOR1 chr15 7220793072209773 11.51 13 + 72208767 chr15 74414585 74415136 7.92 13 chr1581112195 81112934 8.26 16 − 81009005 chr15 82113943 82114802 6.20 20chr15 87755647 87756297 7.33 18 chr16 62621 63380 5.69 14 chr16 49776154978589 6.27 13 + 4948318 chr16 19804086 19804635 8.37 20 chr16 2970373029704362 7.58 15 chr16 31014172 31014616 5.57 13 chr16 49432345 4943301510.49 16 chr16 49744810 49746252 9.90 11 chr16 52873128 52875454 8.32 12− 52874712 52877879 IRX3 chr16 52879121 52880054 7.79 14 chr16 5352334253524273 7.02 16 + 53522611 chr16 53527819 53530073 8.62 11 chr1655229290 55229739 8.87 19 chr16 55234406 55235250 6.28 12 chr16 5525424555254911 7.07 13 chr16 56575924 56577738 6.59 17 + 56567839 chr1665747571 65748520 10.24 14 − 65745589 chr17 1901278 1904018 9.32 12chr17 4385916 4386650 7.11 14 + 4348877 chr17 4635389 4636518 8.20 10 −4635320 4636469 VMO1 − 4635320 chr17 4638694 4639358 7.56 13 chr176500095 6501554 7.20 11 chr17 6866627 6868275 12.02 20 + 6867092 chr176886114 6887363 7.45 14 − 6885672 6887966 SLC16A11 − 6885672 chr1724967181 24969332 12.22 12 − 24965899 chr17 27846829 27847493 7.17 13 −27843740 chr17 32371614 32374986 8.39 13 + 32368611 32374607 LHX1 +32368611 chr17 34584727 34585356 9.69 18 − 34583234 chr17 3693743636938490 8.11 8 chr17 38089183 38089828 5.65 9 + 38088157 chr17 3818926138190408 9.88 13 + 38186221 chr17 41329931 41330876 7.71 22 + 41328944chr17 42251317 42252051 10.35 11 chr17 44151655 44152294 8.85 14 chr1744158959 44159608 8.44 16 − 44157124 chr17 44179727 44180381 9.17 17chr17 44564528 44565791 10.73 10 chr17 45000264 45000898 7.21 11 chr1745396800 45398051 10.87 11 chr17 55570838 55572341 9.24 18 chr1755853774 55854910 6.09 16 chr17 56827847 56829192 7.61 8 chr17 5682985656830915 13.30 14 chr17 69719958 69720487 9.44 11 − 69717735 chr1769781509 69782358 7.08 11 chr17 76724071 76724700 6.36 8 − 76705690chr17 77420013 77420742 6.14 12 − 77418885 chr17 77479685 77480419 7.2413 + 77478995 chr18 5186243 5187292 11.77 12 − 5133671 chr18 2687549426877443 12.24 16 − 26824049 chr18 46048374 46049045 6.75 15 − 4604921346062142 MBD1 chr18 53254565 53259819 10.42 18 + 53253914 chr18 5509192855092557 11.94 15 chr18 72664418 72665346 7.60 10 chr19 335010 3356747.63 11 chr19 554463 555102 6.81 10 + 540892 chr19 1812475 1814434 8.5013 − 1803398 chr19 4279423 4279977 5.17 13 − 4275039 chr19 94697249470804 10.30 11 chr19 10258672 10259721 8.63 13 + 10258649 10260198ICAM4 + 10258649 chr19 10558593 10559142 6.31 12 chr19 11167307 111676565.37 19 − 11135943 chr19 11420829 11421273 5.50 16 + 11407268 chr1913844524 13845198 8.03 18 chr19 18389860 18391505 11.85 15 chr1940222452 40224104 9.89 21 + 40213373 40223193 SCN1B chr19 4040746340408032 4.95 10 − 40407543 40411468 FAM187B chr19 40740718 407415478.85 11 − 40732934 chr19 46008398 46009157 9.53 20 chr19 4633314946333903 9.66 20 chr19 49455836 49456370 7.46 11 chr19 50811149 508118737.00 18 − 50804497 chr19 53516363 53517093 11.19 14 chr19 5360970353610162 5.98 8 + 53589943 chr19 53891560 53892099 7.52 10 + 53891039chr19 54214396 54215050 5.80 11 chr19 54327945 54328592 6.76 12 +54314474 chr19 54628533 54629188 7.58 16 − 54624467 chr19 5463128954632038 8.21 15 − 54624467 chr19 56897971 56898600 8.40 22 + 56888404chr19 59157320 59157774 9.54 19 chr19 60489952 60490616 6.44 19 +60487345 chr19 60721053 60721912 9.92 19 + 60691681 60722277 LOC284297 +60691681 chr2 2752186 2752835 10.69 12 chr2 10100088 10101856 10.60 16chr2 19416426 19417175 7.98 12 − 19414726 chr2 19424458 19425187 8.49 17chr2 72226653 72228187 10.04 12 − 72209874 chr2 74582914 74584780 8.71 8− 74585677 74588329 PCGF1 + 74583251 chr2 86869500 86870049 8.28 11 −86865239 chr2 111591681 111594348 13.30 21 + 111206620 111592270 ACOXLchr2 115636321 115637260 13.24 12 + 115636201 chr2 119322147 11932350210.24 13 chr2 119323782 119324421 11.62 12 chr2 119630604 11963196810.75 17 − 119630288 119632941 C1QL2 − 119630288 chr2 119905436119906957 8.58 13 + 119905915 chr2 128126805 128127459 5.72 8 +128119908 128126682 GPR17 − 128112466 chr2 155263352 155264303 12.69 17chr2 156884504 156885470 9.52 21 chr2 158161857 158162509 7.18 12 −158091524 chr2 160626827 160628085 6.25 14 chr2 171276492 171277161 9.568 − 171277194 171279323 LOC440925 chr2 172659538 172660401 9.52 15 +172658453 chr2 172660882 172661546 8.49 19 + 172658453 chr2 172673432172675478 7.47 13 − 172672411 chr2 176641194 176641858 10.10 15 chr2176653956 176656685 9.31 11 − 176653080 176656936 EVX2 − 176653080 chr2176671999 176672948 7.88 16 + 176672775 176673734 HOXD12 + 176672775chr2 176690151 176690900 10.31 17 + 176689737 chr2 176737592 1767382479.42 16 + 176737050 chr2 200034962 200036119 8.63 15 chr2 200041833200043073 13.73 23 + 200041065 chr2 219481739 219482468 8.63 13 chr2222869621 222872193 7.97 14 + 222871109 chr2 222884739 222885493 8.59 17chr2 232103046 232104293 8.25 15 chr2 232279406 232280745 13.71 9 chr2233442461 233443405 8.93 17 chr2 236751568 236752820 7.95 11 chr203167981 3168945 7.77 17 chr20 21434032 21436910 6.89 8 chr20 2250552122506890 10.23 14 − 22489193 chr20 22515240 22516065 7.90 14 chr2025076779 25077618 6.77 13 chr20 36787544 36790818 19.61 16 + 3678651836791429 SLC32A1 + 36786518 chr20 41251999 41252763 10.05 12 chr2041976531 41978482 10.08 20 + 41976905 chr20 43469538 43470502 10.23 12 +43468650 chr20 43531029 43532764 7.57 11 chr20 43885946 43886595 9.09 20− 43885261 43889360 TNNC2 − 43885261 chr20 44072542 44073411 9.39 19 +44070953 chr20 44073717 44074646 12.54 22 + 44070953 chr20 4411920044121030 7.91 15 + 44091227 chr21 21291294 21291968 10.35 10 chr2134962773 34964708 7.05 10 + 34963557 chr21 36990087 36992825 11.85 15chr22 28039290 28041531 13.41 21 − 28038921 28041748 RASL10A − 28038921chr22 36807805 36808832 5.62 12 − 36804089 chr22 38114321 38115070 6.5718 chr3 2115949 2117089 11.08 19 chr3 32254374 32255424 7.95 11 chr350357760 50358604 8.66 9 chr3 131176854 131177513 8.41 14 + 131175803chr3 138020672 138021936 18.36 10 + 138020550 chr3 155629021 15562967010.35 10 − 155538154 chr3 159303324 159304476 14.83 12 − 159296493 chr3161649899 161651150 6.32 16 chr3 173648073 173649732 9.01 18 chr3188870393 188870937 13.49 19 chr4 560951 561800 9.51 11 chr4 13973931398032 7.81 13 chr4 2903227 2903861 7.18 10 − 2902086 chr4 49054834906134 5.98 14 chr4 5945893 5946532 7.96 15 chr4 13134417 13135173 7.9415 chr4 13135429 13136158 6.92 13 chr4 30332624 30333968 9.09 19 +30331134 chr4 41443929 41444657 8.23 16 − 41440855 chr4 4157004741570684 10.05 23 chr4 41577137 41577766 10.06 15 chr4 52612173 526130187.73 17 chr4 54661877 54662826 14.11 17 + 54661004 54662879 GSX2 +54661004 chr4 85623804 85624451 7.86 11 chr4 85636686 85637940 7.21 15 −85633459 chr4 94974738 94975381 8.87 9 chr4 111758660 111759824 11.15 19− 111758028 111777957 PITX2 − 111758028 chr4 111761532 111764385 6.14 11− 111758028 chr4 122211474 122212323 18.69 9 − 122176231 chr4 154932988154933738 10.82 17 chr4 174664270 174664919 9.49 14 chr4 174674369174675025 8.92 16 chr4 174684839 174685488 6.03 12 − 174684226 174687953HAND2 − 174684226 chr4 174688523 174689572 9.32 15 + 174688183 chr52792454 2794206 7.35 10 chr5 2805770 2806724 9.19 18 + 2805261 chr53643658 3644997 8.79 15 chr5 32747979 32750229 12.42 16 + 32747421 chr537872516 37875378 15.39 13 − 37851509 chr5 43027793 43028447 15.00 8chr5 43053743 43054382 16.74 9 chr5 54554757 54555401 8.65 11 chr563292326 63293075 10.85 20 chr5 76151227 76151876 8.32 17 + 76150588chr5 76968098 76969322 7.73 9 − 76960292 chr5 77176139 77176798 13.09 12chr5 140286488 140287347 15.62 17 + 140242037 chr5 140742560 1407431198.07 13 + 140733834 chr5 140747458 140748016 8.51 17 + 140742650 chr5140767256 140768329 11.36 19 + 140762703 chr5 145698688 145700318 10.4612 + 145698779 145700276 POU4F3 + 145698779 chr5 154006904 15400754311.15 12 chr5 157030866 157031611 9.76 19 chr5 158456493 158457132 12.4025 − 158055500 chr5 158463748 158464392 9.38 15 chr5 158690162 1586915367.47 12 chr5 172596242 172597176 10.35 17 chr5 172604911 172605563 6.2613 chr6 1550805 1554054 7.29 8 chr6 5939707 5940356 6.99 18 chr610497957 10499310 8.37 15 chr6 10530143 10530792 6.97 11 chr6 1099497210995711 7.30 13 chr6 26125798 26126758 9.25 13 chr6 27215069 272156336.95 16 + 27215066 27215436 HIST1H4I − 27214051 chr6 27387778 2738864713.86 15 chr6 27948718 27949880 10.00 11 − 27948904 27949268 HIST1H4L −27948904 chr6 28283085 28283729 13.91 13 chr6 29063972 29064638 9.18 14chr6 30050074 30050923 5.05 13 chr6 30818753 30820599 10.08 16 −30818954 30820306 IER3 − 30818954 chr6 37611537 37612266 7.91 10 chr639977327 39978066 7.40 15 + 39868770 chr6 41447224 41447987 15.75 14chr6 41449448 41450517 15.53 10 chr6 41623448 41624191 15.17 21 +41622141 chr6 42253599 42254248 12.27 14 + 42231121 chr6 4372000043721360 8.21 12 chr6 78228956 78230415 10.96 17 chr6 83130068 831312088.72 14 chr6 85539298 85540860 9.21 12 chr6 100173150 100173899 9.05 9chr6 101009995 101010637 15.23 15 − 100943470 chr6 101011950 1010134476.65 16 − 100943470 chr6 112147783 112148610 9.95 13 − 112089177 chr6117192435 117193290 8.00 14 − 117180053 chr6 137857127 137857980 13.2214 chr6 146797042 146797694 10.61 14 + 146390474 chr6 150326335150328184 32.01 19 + 150326835 chr7 526958 527907 9.01 15 chr7 50777795078727 6.16 15 + 5078263 5079380 LOC389458 + 5078263 chr7 84485058449259 14.36 21 + 8440109 chr7 8449600 8450254 10.16 21 + 8440109 chr725864688 25865353 11.40 14 chr7 27093668 27094904 10.45 13 chr7 2710057027102300 8.47 12 − 27099138 chr7 27156749 27161918 7.67 13 − 2715986227162821 HOXA7 − 27159862 chr7 27177747 27179316 8.87 11 − 27176734 chr727185469 27186510 12.70 12 − 27187300 27191360 HOXA11 chr7 2719064827192605 14.02 16 + 27191551 chr7 27193961 27196205 9.17 15 + 2719155127195437 HOXA11AS + 27191551 chr7 27210155 27212509 8.41 11 chr727215870 27217214 10.22 9 chr7 27241669 27242703 6.76 12 chr7 2724865727253218 8.83 9 + 27248688 27252717 EVX1 + 27248688 chr7 2725746927259401 8.39 19 chr7 44119525 44120164 6.06 15 − 44120810 44129672POLD2 + 44110484 chr7 44151320 44152244 7.95 18 − 44150394 44195547 GCK− 44150394 chr7 51352205 51352779 7.08 20 chr7 63667353 63668297 6.38 12chr7 96488377 96489524 7.61 11 − 96487637 96492079 DLX5 − 96487637 chr7101722805 101723452 5.94 15 + 101715165 chr7 107429031 107429905 11.0216 − 107351481 chr7 116906858 116907422 10.33 12 chr7 120755525120756474 10.97 17 + 120752656 chr7 127595227 127596086 10.92 12 chr7127697971 127698610 6.31 16 chr7 129207840 129209778 8.11 15 chr7129210064 129210736 8.06 13 chr7 129912506 129914030 8.21 18 − 129914133129918249 MESTIT1 chr7 131983416 131984045 7.46 10 chr7 148743033148743764 17.18 22 chr7 152252636 152253475 11.38 26 chr7 154857518154859486 8.64 14 chr7 154951604 154952844 6.66 12 chr7 155291555155292209 7.84 17 − 155288318 chr7 156125196 156126661 8.12 9 chr7158515940 158517089 7.67 11 − 158513626 chr8 206253 206907 5.98 14 chr810626015 10628284 7.94 10 chr8 11574478 11576337 7.53 14 chr8 2361842223620876 8.71 14 − 23615909 chr8 24826911 24828465 8.26 19 + 24827178chr8 24868882 24870630 9.36 13 − 24864387 chr8 25965058 25965627 10.3017 chr8 26777575 26780521 15.03 22 − 26683138 chr8 41543514 415443537.01 14 chr8 49471563 49472223 8.17 8 chr8 57522214 57523567 8.06 16chr8 65873955 65874718 9.27 15 chr8 71144440 71147178 6.16 11 − 71126576chr8 95720330 95720979 6.32 19 chr8 97238932 97239559 6.75 16 − 97223733chr8 98359200 98360130 6.68 12 chr8 100029975 100030644 10.56 13 +100025806 chr8 102573379 102574213 9.31 19 chr8 110725080 110726354 9.5513 − 110655580 chr8 145896215 145896964 8.22 19 chr9 34369432 343712817.87 20 − 34369016 34387849 C9orf24 − 34369016 chr9 78818627 7882011613.23 11 chr9 85760867 85761816 9.92 8 chr9 90339041 90340305 7.18 17chr9 99109616 99110142 9.01 14 + 99040599 chr9 99304529 99305158 9.93 11chr9 124021380 124022605 12.53 13 − 124004678 chr9 125810809 12581194013.91 12 chr9 129501281 129501822 7.67 12 chr9 135557390 135558039 7.4115 − 135518506 chrX 111211507 111212571 11.31 13 − 110904197 chrY 510532512001 5.99 11 + 505078 Peaks Gene Upstream Gene chr Start End EndSymbol Strand Start End Symbol chr1 2456587 2457246 chr1 3797329737974046 38003411 EPHA10 chr1 38002438 38003587 − 37998531 38003411EPHA10 chr1 46712589 46713323 chr1 47469314 47470340 chr1 5459442054595152 54644680 SSBP3 chr1 60311689 60312524 − 60228653 60312014C1orf87 chr1 63556597 63560345 + 63561317 63563385 FOXD3 chr1 9096329090965440 chr1 111614814 111615476 chr1 119328183 119328832 119333702TBX15 chr1 143012194 143012944 143053112 LOC728875 chr1 144674586144675239 chr1 144690585 144691329 chr1 144702031 144702576 chr1145971587 145972240 chr1 145987585 145988329 chr1 145999042 145999587chr1 146184793 146185549 chr1 146219303 146219957 chr1 146248547146249292 chr1 146256160 146256719 chr1 146466570 146467421 chr1147599481 147600124 chr1 147600226 147600785 chr1 147665315 147665969chr1 147882409 147883136 chr1 147985011 147986318 chr1 150078663150079332 150083264 LOC100132111 − 150076962 150079657 LOC100191040 chr1150349668 150350912 150353180 TCHH chr1 153320294 153321238 153326638EFNA3 chr1 153557083 153557827 − 153557263 153567533 RUSC1 chr1154527610 154528254 154528858 TMEM79 chr1 155085184 155085851 155095290INSRR chr1 165119820 165120544 chr1 179719350 179719904 + 179719338180037338 CACNA1E chr1 181421423 181422187 + 181421796 181480885 LAMC2chr1 196156945 196157494 196165896 LHX9 chr1 207914587 207915953207916358 G0S2 + 207915292 207916358 G0S2 chr1 219131519 219132268 chr1219134095 219134834 chr1 222002983 222003622 222030343 CAPN2 chr1222870205 222872650 222994872 CNIH3 + 222870801 222994872 CNIH3 chr1226700008 226700767 chr1 234752777 234754328 234779619 LGALS8 +234753699 234779619 LGALS8 chr10 7489395 7490153 7493456 SFMBT2 chr1046412477 46413703 + 46413551 46420574 GPRIN2 chr10 50486546 50487915 +50488352 50490772 SLC18A3 chr10 50488946 50491715 50490772 SLC18A3 +50492088 50543156 CHAT chr10 79066119 79066773 79067583 KCNMA1 −78314641 79067583 KCNMA1 chr10 81732969 81733640 chr10 99070134 9907106399071662 FRAT1 chr10 102580005 102580574 chr10 102972304 102973663 chr10120344105 120345734 120345150 PRLHR − 120342905 120345150 PRLHR chr10124628733 124629607 + 124629138 124648220 LOC399815 chr11 17189921720660 1742077 HCCA2 chr11 1721203 1723357 chr11 1727398 1728562 chr112102906 2103345 chr11 2183194 2184458 chr11 10428000 10428654 + 1042879910485702 AMPD3 chr11 14883948 14884302 chr11 19220281 19221020 chr1122319354 22320095 22357620 SLC17A6 chr11 31789500 31795000 31789361 PAX6− 31794625 31865159 DKFZp686K1684 chr11 32411453 32412722 32413663 WT1 −32365900 32413663 WT1 chr11 44283814 44284478 44288292 ALX4 chr1144935052 44935706 chr11 63531706 63532455 63690161 MACROD1 chr1163823533 63825290 + 63824438 63828815 C11orf20 chr11 64947186 64947820chr11 73049464 73050318 73051512 PLEKHB1 chr11 94523361 94524500 chr11124133793 124135432 124137433 ESAM chr11 133762389 133763138 133787022B3GAT1 chr12 6803351 6804025 6806844 GPR162 chr12 21985422 21986671chr12 42231009 42232738 − 42034278 42231991 ADAMTS20 chr12 4551120145511951 chr12 47771091 47771619 47774869 DHH chr12 47975997 4797764647978748 PRPH + 47975175 47978748 PRPH chr12 52418168 52418832 chr1252646454 52647108 52648782 HOTAIR chr12 53049097 53049742 chr12 5516827155169242 − 55151002 55168448 GLS2 chr12 55918204 55919848 55920742NDUFA4L2 − 55914952 55920742 NDUFA4L2 chr12 56300983 56301939 56306201SLC26A10 chr12 79626169 79626805 79627382 MYF6 + 79625576 79627382 MYF6chr12 84197229 84198765 + 84198166 84219692 ALX1 chr12 101868310101868969 chr12 113322635 113323304 113326086 TBX5 chr12 115800795115801644 115803615 HRK chr13 27399628 27401487 chr13 4092892540929564 + 40929541 40943013 C13orf15 chr13 52211145 52212607 − 5217539952211948 LECT1 chr13 102224101 102224965 − 102216465 102224150 C13orf27chr13 112725390 112726034 112800863 MCF2L chr14 22360909 22361365 chr1423115638 23116287 23117849 JPH4 chr14 36061370 36062206 chr14 3612220936123483 − 36118966 36121537 NKX2-8 chr14 36194850 36195499 chr1436196539 36197988 36216762 PAX9 + 36196532 36216762 PAX9 chr14 3620153936202583 36216762 PAX9 chr14 37137508 37138866 chr14 37748210 3775057037752019 SSTR1 chr14 53488199 53488653 53493304 BMP4 chr14 5349163653492707 53493304 BMP4 − 53486204 53493304 BMP4 chr14 53499835 53500364chr14 56344682 56346630 56346937 OTX2 − 56337177 56346937 OTX2 chr1460021741 60022400 − 59972426 60022517 C14orf39 chr14 60043368 60044022chr14 60045099 60047942 60048278 SIX6 + 60045690 60048278 SIX6 chr1473776450 73777601 73799194 VSX2 + 73775927 73799194 VSX2 chr14 7414708374147922 74148787 LTBP2 − 74034638 74148787 LTBP2 chr14 9430439094305319 94306252 GSC − 94304312 94306252 GSC chr14 94308930 94310289chr14 105023673 105024599 + 105024301 105026169 CRIP1 chr15 4319586943196818 43197593 DUOXA2 chr15 43783619 43784273 chr15 50867775 5086892450869501 ONECUT1 − 50836644 50869501 ONECUT1 chr15 50873886 50875041chr15 62984637 62985202 chr15 65913980 65915431 chr15 72207930 7220977372216196 ISLR2 + 72209795 72216196 ISLR2 chr15 74414585 74415136 chr1581112195 81112934 81113783 CPEB1 − 81009005 81113783 CPEB1 chr1582113943 82114802 + 82113841 82499597 ADAMTSL3 chr15 87755647 87756297chr16 62621 63380 − 48057 62629 RHBDF1 chr16 4977615 4978589 5009157SEC14L5 chr16 19804086 19804635 − 19777793 19803652 GPRC5B chr1629703730 29704362 chr16 31014172 31014616 − 31009675 31013777 VKORC1chr16 49432345 49433015 chr16 49744810 49746252 chr16 52873128 52875454chr16 52879121 52880054 chr16 53523342 53524273 53525896 IRX5 + 5352261153525896 IRX5 chr16 53527819 53530073 chr16 55229290 55229739 + 5523007855231500 MT1A chr16 55234406 55235250 + 55235216 55236353 MT1DP chr1655254245 55254911 chr16 56575924 56577738 56579518 TEPP chr16 6574757165748520 65751313 TRADD chr17 1901278 1904018 chr17 4385916 43866504389977 SPNS2 chr17 4635389 4636518 4636469 VMO1 − 4635320 4636469 VMO1chr17 4638694 4639358 + 4638993 4640425 GLTPD2 chr17 6500095 6501554chr17 6866627 6868275 6873685 BCL6B + 6867092 6873685 BCL6B chr176886114 6887363 6887966 SLC16A11 − 6885672 6887966 SLC16A11 chr1724967181 24969332 24972620 CORO6 chr17 27846829 27847493 28228015 MYO1Dchr17 32371614 32374986 32374607 LHX1 chr17 34584727 34585356 34607427CACNB1 chr17 36937436 36938490 − 36933394 36938167 KRT19 chr17 3808918338089828 38105536 CNTNAP1 chr17 38189261 38190408 38202610 WNK4 chr1741329931 41330876 41331959 LOC100130148 + 41328944 41331959 LOC100130148chr17 42251317 42252051 − 42196856 42251081 WNT3 chr17 44151655 44152294chr17 44158959 44159608 44161110 HOXB13 chr17 44179727 44180381 chr1744564528 44565791 + 44565327 44602350 B4GALNT2 chr17 45000264 45000898chr17 45396800 45398051 chr17 55570838 55572341 chr17 5585377455854910 + 55854646 55863568 C17orf64 chr17 56827847 56829192 chr1756829856 56830915 chr17 69719958 69720487 69721055 MGC16275 + 6972129069769752 TTYH2 chr17 69781509 69782358 + 69781991 69822617 DNAI2 chr1776724071 76724700 76754467 AATK chr17 77420013 77420742 77422527 ARHGDIAchr17 77479685 77480419 77481919 LOC92659 + 77478995 77481919 LOC92659chr18 5186243 5187292 5187255 LOC642597 − 5133671 5187255 LOC642597chr18 26875494 26877443 26876779 DSC3 − 26824049 26876779 DSC3 chr1846048374 46049045 chr18 53254565 53259819 53309528 ONECUT2 + 5325391453309528 ONECUT2 chr18 55091928 55092557 − 55085246 55091605 RAX chr1872664418 72665346 + 72665103 72811670 ZNF236 chr19 335010 335674 chr19554463 555102 568159 HCN2 chr19 1812475 1814434 1814564 KLF16 − 18033981814564 KLF16 chr19 4279423 4279977 4289847 STAP2 chr19 9469724 9470804− 9438030 9470279 ZNF560 chr19 10258672 10259721 10260198 ICAM4 +10258649 10260198 ICAM4 chr19 10558593 10559142 − 10544347 10558991AP1M2 chr19 11167307 11167656 11169224 KANK2 chr19 11420829 1142127311422782 PRKCSH chr19 13844524 13845198 chr19 18389860 18391505 +18391135 18406459 SSBP4 chr19 40222452 40224104 + 40223249 40249315 HPNchr19 40407463 40408032 chr19 40740718 40741547 40746400 ATP4A chr1946008398 46009157 chr19 46333149 46333903 chr19 49455836 49456370 +49455915 49471308 ZNF233 chr19 50811149 50811873 50834509 EML2 chr1953516363 53517093 chr19 53609703 53610162 53640000 GRIN2D chr19 5389156053892099 53901003 FUT2 + 53891039 53901003 FUT2 chr19 54214396 54215050chr19 54327945 54328592 54346092 PPFIA3 chr19 54628533 54629188 54636620SLC17A7 chr19 54631289 54632038 54636620 SLC17A7 chr19 56897971 5689860056900254 NCRNA00085 chr19 59157320 59157774 + 59158105 59177951 CACNG8chr19 60489952 60490616 60515715 BRSK1 chr19 60721053 60721912 60722277LOC284297 chr2 2752186 2752835 chr2 10100088 10101856 + 1010113210112414 KLF11 chr2 19416426 19417175 19421853 OSR1 chr2 1942445819425187 chr2 72226653 72228187 72228471 CYP26B1 − 72209874 72228471CYP26B1 chr2 74582914 74584780 74585700 LOC151534 + 74583251 74585700LOC151534 chr2 86869500 86870049 86889030 CD8A chr2 111591681111594348 + 111594961 111642493 BCL2L11 chr2 115636321 115637260116318406 DPP10 + 115636201 116318406 DPP10 chr2 119322147 119323502 −119316216 119322229 EN1 chr2 119323782 119324421 chr2 119630604119631968 119632941 C1QL2 − 119630288 119632941 C1QL2 chr2 119905436119906957 119912566 TMEM37 + 119905915 119912566 TMEM37 chr2 128126805128127459 128155830 LIMS2 chr2 155263352 155264303 + 155263338 155421260KCNJ3 chr2 156884504 156885470 chr2 158161857 158162509 158193645 ACVR1C− 158091524 158162318 ACVR1C chr2 160626827 160628085 − 160510571160627367 PLA2R1 chr2 171276492 171277161 chr2 172659538 172660401172662647 DLX1 chr2 172660882 172661546 172662647 DLX1 chr2 172673432172675478 172675724 DLX2 − 172672411 172675724 DLX2 chr2 176641194176641858 chr2 176653956 176656685 176656936 EVX2 − 176653080 176656936EVX2 chr2 176671999 176672948 176673734 HOXD12 + 176672775 176673734HOXD12 chr2 176690151 176690900 176692916 HOXD10 + 176689737 176692916HOXD10 chr2 176737592 176738247 176746072 HOXD3 + 176737050 176746072HOXD3 chr2 200034962 200036119 chr2 200041833 200043073 200045724FLJ32063 + 200041065 200045724 FLJ32063 chr2 219481739 219482468 chr2222869621 222872193 222878180 CCDC140 + 222871109 222878180 CCDC140 chr2222884739 222885493 chr2 232103046 232104293 − 232096114 232103426 NMUR1chr2 232279406 232280745 + 232281478 232286494 PTMA chr2 233442461233443405 + 233443237 233449351 UNQ830 chr2 236751568 236752820 chr203167981 3168945 chr20 21434032 21436910 chr20 22505521 22506890 22507280C20orf56 − 22489193 22507280 C20orf56 chr20 22515240 22516065 chr2025076779 25077618 − 25069433 25077426 LOC284798 chr20 36787544 3679081836791429 SLC32A1 chr20 41251999 41252763 − 40134805 41251971 PTPRT chr2041976531 41978482 42131668 TOX2 + 41978195 42131668 TOX2 chr20 4346953843470502 43472661 DBNDD2 + 43470328 43472661 DBNDD2 chr20 4353102943532764 + 43531807 43543586 WFDC2 chr20 43885946 43886595 43889360TNNC2 chr20 44072542 44073411 44078607 MMP9 chr20 44073717 4407464644078607 MMP9 chr20 44119200 44121030 44122196 SLC12A5 chr21 2129129421291968 + 21292503 21833085 NCAM2 chr21 34962773 34964708 35012389CLIC6 + 34963557 35012389 CLIC6 chr21 36990087 36992825 chr22 2803929028041531 28041748 RASL10A − 28038921 28041748 RASL10A chr22 3680780536808832 36809116 SLC16A8 − 36804089 36809116 SLC16A8 chr22 3811432138115070 chr3 2115949 2117089 + 2117246 3074645 CNTN4 chr3 3225437432255424 + 32255174 32386817 CMTM8 chr3 50357760 50358604 − 5035354050358160 ZMYND10 chr3 131176854 131177513 131179466 TRH chr3 138020672138021936 138057424 TMEM22 + 138020550 138057424 TMEM22 chr3 155629021155629670 155630198 GPR149 − 155538154 155630198 GPR149 chr3 159303324159304476 159306630 SHOX2 − 159296493 159303520 SHOX2 chr3 161649899161651150 − 161635984 161650320 TRIM59 chr3 173648073 173649732 −173648027 173648897 GHSR chr3 188870393 188870937 − 188869387 188870895SST chr4 560951 561800 chr4 1397393 1398032 chr4 2903227 2903861 2906384MFSD10 chr4 4905483 4906134 chr4 5945893 5946532 − 5873391 5945686 CRMP1chr4 13134417 13135173 chr4 13135429 13136158 chr4 30332624 3033396830757519 PCDH7 chr4 41443929 41444657 41445744 PHOX2B chr4 4157004741570684 chr4 41577137 41577766 chr4 52612173 52613018 + 5261234952658215 SPATA18 chr4 54661877 54662826 54662879 GSX2 + 5466100454662879 GSX2 chr4 85623804 85624451 chr4 85636686 85637940 85638411NKX6-1 − 85633459 85638411 NKX6-1 chr4 94974738 94975381 chr4 111758660111759824 111777957 PITX2 chr4 111761532 111764385 111777957 PITX2 chr4122211474 122212323 122213123 C4orf31 − 122176231 122213123 C4orf31 chr4154932988 154933738 chr4 174664270 174664919 chr4 174674369 174675025chr4 174684839 174685488 174687953 HAND2 chr4 174688523 174689572174699556 NBLA00301 + 174688183 174699556 NBLA00301 chr5 2792454 2794206chr5 2805770 2806724 2808511 C5orf38 + 2805261 2808511 C5orf38 chr53643658 3644997 chr5 32747979 32750229 32823011 NPR3 + 32747421 32823011NPR3 chr5 37872516 37875378 37875539 GDNF − 37851509 37875539 GDNF chr543027793 43028447 chr5 43053743 43054382 chr5 54554757 54555401 chr563292326 63293075 − 63292033 63293302 HTR1A chr5 76151227 7615187676166895 F2RL1 + 76150588 76166895 F2RL1 chr5 76968098 76969322 76970278OTP − 76960292 76970278 OTP chr5 77176139 77176798 chr5 140286488140287347 140372113 PCDHA13 + 140286485 140372113 PCDHAC1 chr5 140742560140743119 140872730 PCDHGA6 + 140742650 140872730 PCDHGA7 chr5 140747458140748016 140872730 PCDHGA7 + 140747635 140872730 PCDHGB4 chr5 140767256140768329 140872730 PCDHGA9 + 140767953 140872730 PCDHGB6 chr5 145698688145700318 145700276 POU4F3 + 145698779 145700276 POU4F3 chr5 154006904154007543 chr5 157030866 157031611 + 157031138 157039738 C5orf52 chr5158456493 158457132 158459366 EBF1 chr5 158463748 158464392 chr5158690162 158691536 − 158674368 158690059 IL12B chr5 172596242 172597176chr5 172604911 172605563 chr6 1550805 1554054 chr6 5939707 5940356 chr610497957 10499310 chr6 10530143 10530792 chr6 10994972 10995711 +10995049 11082527 SYCP2L chr6 26125798 26126758 − 26125238 26126019HIST1H1A chr6 27215069 27215633 27222598 HIST1H2BK chr6 2738777827388647 chr6 27948718 27949880 27949268 HIST1H4L − 27948904 27949268HIST1H4L chr6 28283085 28283729 chr6 29063972 29064638 chr6 3005007430050923 + 30050870 30054156 HCG9 chr6 30818753 30820599 30820306 IER3 −30818954 30820306 IER3 chr6 37611537 37612266 chr6 39977327 3997806639980619 DAAM2 chr6 41447224 41447987 chr6 41449448 41450517 chr641623448 41624191 41678100 FOXP4 chr6 42253599 42254248 42255770 GUCA1Achr6 43720000 43721360 + 43720787 43746722 RSPH9 chr6 78228956 78230415− 78228666 78229839 HTR1B chr6 83130068 83131208 + 83130257 83133335TPBG chr6 85539298 85540860 chr6 100173150 100173899 chr6 101009995101010637 101018272 SIM1 chr6 101011950 101013447 101018272 SIM1 chr6112147783 112148610 112301320 FYN − 112089177 112147958 FYN chr6117192435 117193290 117193579 FAM162B − 117180053 117193579 FAM162B chr6137857127 137857980 − 137855028 137857224 OLIG3 chr6 146797042 146797694146800424 GRM1 chr6 150326335 150328184 150336539 ULBP1 + 150326835150336539 ULBP1 chr7 526958 527907 − 503424 526007 PDGFA chr7 50777795078727 5079380 LOC389458 + 5078263 5079380 LOC389458 chr7 84485058449259 8759118 NXPH1 chr7 8449600 8450254 8759118 NXPH1 chr7 2586468825865353 chr7 27093668 27094904 chr7 27100570 27102300 27102150 HOXA1 −27099138 27102150 HOXA1 chr7 27156749 27161918 27162821 HOXA7 − 2715986227162821 HOXA7 chr7 27177747 27179316 27186401 HOXA10 chr7 2718546927186510 − 27176734 27186401 HOXA10 chr7 27190648 27192605 27195437HOXA11AS + 27191551 27195437 HOXA11AS chr7 27193961 27196205 27195437HOXA11AS chr7 27210155 27212509 chr7 27215870 27217214 chr7 2724166927242703 chr7 27248657 27253218 27252717 EVX1 + 27248688 27252717 EVX1chr7 27257469 27259401 chr7 44119525 44120164 44120684 AEBP1 chr744151320 44152244 44195547 GCK chr7 51352205 51352779 − 5105140251352009 COBL chr7 63667353 63668297 chr7 96488377 96489524 96492079DLX5 chr7 101722805 101723452 101748898 SH2B2 chr7 107429031 107429905107431040 LAMB1 chr7 116906858 116907422 + 116907252 117095954 CFTR chr7120755525 120756474 120768394 WNT16 + 120756325 120768394 WNT16 chr7127595227 127596086 chr7 127697971 127698610 chr7 129207840 129209778chr7 129210064 129210736 chr7 129912506 129914030 + 129913281 129933367MEST chr7 131983416 131984045 − 131820060 131983987 PLXNA4 chr7148743033 148743764 chr7 152252636 152253475 chr7 154857518 154859486chr7 154951604 154952844 chr7 155291555 155292209 155297728 SHH chr7156125196 156126661 + 156126161 156162581 RNF32 chr7 158515940 158517089158630410 VIPR2 chr8 206253 206907 chr8 10626015 10628284 − 1061868710625432 SOX7 chr8 11574478 11576337 chr8 23618422 23620876 23619867NKX2-6 − 23615909 23619867 NKX2-6 chr8 24826911 24828465 24832511 NEFM +24828359 24832511 NEFM chr8 24868882 24870630 24870048 NEFL − 2486438724870048 NEFL chr8 25965058 25965627 chr8 26777575 26780521 26778839ADRA1A − 26683138 26778839 ADRA1A chr8 41543514 41544353 chr8 4947156349472223 chr8 57522214 57523567 − 57516068 57521836 PENK chr8 6587395565874718 − 65671082 65873902 CYP7B1 chr8 71144440 71147178 71146116PRDM14 − 71126576 71146116 PRDM14 chr8 95720330 95720979 chr8 9723893297239559 97242196 GDF6 chr8 98359200 98360130 − 98354889 98359352 TSPYL5chr8 100029975 100030644 100033500 OSR2 chr8 102573379 102574213 +102573843 102751128 GRHL2 chr8 110725080 110726354 110773196 GOLSYN −110655580 110726998 GOLSYN chr8 145896215 145896964 chr9 3436943234371281 34387849 C9orf24 − 34369016 34371598 C9orf24 chr9 7881862778820116 chr9 85760867 85761816 − 85743046 85761483 C9orf64 chr990339041 90340305 + 90339835 90380523 NXNL2 chr9 99109616 9911014299179390 KIAA1529 chr9 99304529 99305158 chr9 124021380 124022605124030840 LHX6 chr9 125810809 125811940 chr9 129501281 129501822 chr9135557390 135558039 135594898 SARDH chrX 111211507 111212571 111212660TRPC5 + 111212908 111587129 ZCCHC16 chrY 510532 512001 540146 SHOX *This column shows the Average P value score of the peak in all tumors,excluding the tumors that do not have this peak.

TABLE 2 Frequently methylated regions in grade I tumors No. of grade ISeq. ID tumors Adjacent NO. Chr Start End methylated gene 2 chr5158456493 158457132 6 EBF1 11 chr1 2456587 2457246 5 12 chr1 147665315147665969 5 13 chr11 1718992 1720660 5 HCCA2 14 chr11 44935052 449357065 15 chr17 4385916 4386650 5 16 chr19 554463 555102 5 HCN2 17 chr744119525 44120164 5 POLD2

TABLE 3 Functional annotation of 5′-end methylated genes in tumorsamples P GO ID GO Term Count Value Benjamini Genes GO: 0048513 organ 347.50E−09 1.33E−05 IL12B, PDGFA, LECT1, BMP4, OTX2, development MYF6,CNTN4, PAX9, FYN, SHOX2, EN1, CHAT, GSX2, KRT19, SIX6, POU4F3, RAX,FOXD3, GDNF, HOXD12, NPR3, HOXD3, HOXD10, MEST, ONECUT2, ALX1, NKX2-6,ONECUT1, OTP, GHSR, LAMC2, DLX2, NKX6-1, CFTR GO: 0007420 brain 111.31E−06 1.16E−03 CNTN4, ALX1, RAX, OTP, FYN, EN1, development DLX2,BMP4, GSX2, OTX2, CFTR GO: 0007399 nervous system 22 3.38E−06 1.99E−03POU4F3, RAX, NCAM2, GDNF, BMP4, development OTX2, NEFM, PCDHAC1, CNTN4,ALX1, OTP, FYN, GHSR, EN1, SHOX2, DLX2, CHAT, NKX6-1, GSX2, CRMP1,NKX2-8, CFTR GO: 0031323 regulation of 47 8.01E−06 3.54E−03 ZNF560,ZNF233, EVX2, IL12B, PDGFA, cellular BMP4, VSX2, OTX2, KLF11, MYF6,metabolic IRX5, PAX9, GSC, KLF16, EN1, SHOX2, process GSX2, GRHL2, SOX7,SIX6, NKX2-8, CPEB1, TOX2, POU4F3, RAX, FOXD3, BCL6B, HOXD12, HOXD3,HOXD10, ZNF236, ONECUT2, ALX1, C14orf39, EVX1, HOXA1, ONECUT1, NKX2-6,OLIG3, OTP, PRDM14, GHSR, DLX2, NKX6-1, HOXA10, WT1, HOXA7 GO: 0022008neurogenesis 13 1.49E−05 1.28E−02 POU4F3, GDNF, OTX2, NEFM, CNTN4, OTP,FYN, EN1, CHAT, GSX2, NKX6-1, NKX2-8, CFTR GO: 0048699 generation of 124.40E−05 1.29E−02 CNTN4, POU4F3, OTP, GDNF, FYN, neurons EN1, CHAT,GSX2, OTX2, NEFM, NKX2- 8, CFTR GO: 0045449 regulation of 42 4.73E−051.19E−02 ZNF560, ZNF233, EVX2, VSX2, OTX2, transcription KLF11, MYF6,IRX5, PAX9, GSC, KLF16, EN1, SHOX2, GSX2, GRHL2, SOX7, NKX2-8, SIX6,TOX2, POU4F3, RAX, FOXD3, BCL6B, HOXD12, HOXD3, HOXD10, ZNF236, ONECUT2,ALX1, C14orf39, EVX1, HOXA1, ONECUT1, NKX2-6, OLIG3, OTP, PRDM14, DLX2,NKX6-1, HOXA10, WT1, HOXA7

TABLE 4 Genes methylated at the 5′ end in normal brain Average P P valuescores chr start end accession gene_symbol value score N1 N2 N3 N4 N5 N6chr1 886785 890940 NM_032129 PLEKHN1 14.97 14.73 14.26 18.95 16.13 13.3912.36 chr1 1256597 1258841 NM_152228 TAS1R3 7.66 10.17 8.94 7.32 6.497.39 5.65 chr1 1557590 1559635 NR_002946 MMP23A 9.95 5.89 10.87 12.4712.45 11.30 6.72 chr1 1848895 1850056 NM_001003808 C1orf222 13.14 21.4411.07 16.57 15.68 6.68 7.41 chr1 1994170 1995508 NM_001033581 PRKCZ 6.977.88 6.84 8.28 5.99 6.77 6.03 chr1 6008276 6009325 NM_172130 KCNAB2 8.4110.52 8.43 6.98 11.06 7.36 6.09 chr1 25163901 25164846 NM_001031680RUNX3 10.59 14.26 12.54 9.43 10.79 6.05 10.45 chr1 27062120 27062979NM_006142 SFN 8.59 9.93 7.74 8.18 7.46 11.59 6.63 chr1 34998589 35000128NM_153212 GJB4 32.79 28.52 38.88 41.56 33.17 34.92 19.66 chr1 4586080245861741 NM_001114938 CCDC17 11.04 14.19 8.53 10.48 11.09 13.45 8.47chr1 53696980 53698808 NM_033067 DMRTB1 16.60 8.16 9.53 17.01 19.6721.62 23.62 chr1 55043525 55044789 NM_152607 C1orf177 9.92 11.63 9.5411.13 12.03 9.28 5.93 chr1 64441283 64442237 NM_152489 UBE2U 8.66 8.517.14 10.45 11.00 6.21 8.62 chr1 68288592 68290258 NM_004675 DIRAS3 34.8646.15 25.24 43.22 27.90 39.78 26.86 chr1 92186921 92187877 NM_207189BRDT 15.43 17.69 9.18 15.99 18.51 18.79 12.40 chr1 92455965 92456520NM_001012425 C1orf146 12.21 12.28 13.35 11.96 10.45 12.41 12.80 chr1159761270 159762009 NM_002155 HSPA6 9.99 10.87 8.67 10.34 14.67 8.666.72 chr1 165224549 165225393 NM_032858 MAEL 13.73 12.09 11.89 16.1714.70 12.71 14.80 chr1 221634128 221634757 NM_152610 C1orf65 10.11 14.528.17 12.32 9.59 7.25 8.82 chr1 226069650 226070909 NM_183062 MPN2 8.129.28 8.01 8.18 10.52 6.96 5.76 chr1 226679129 226679963 NM_003493HIST3H3 24.80 30.55 24.80 26.21 22.78 26.66 17.81 chr1 245681115245682749 NM_001004492 OR2B11 12.58 17.09 10.95 13.64 17.28 8.10 8.39chr1 246166331 246167265 NM_175911 OR2L13 9.62 13.82 8.69 7.55 9.67 8.379.61 chr10 84545 86000 NM_177987 TUBB8 26.76 23.45 20.43 27.58 24.0130.78 34.34 chr10 1194985 1196034 NR_015376 C10orf139 35.51 31.12 35.4247.77 38.90 32.47 27.35 chr10 5396191 5397142 NM_053049 UCN3 8.37 11.046.07 7.89 11.89 7.39 5.91 chr10 5530786 5532355 NM_017422 CALML5 26.6234.88 22.07 30.92 28.60 27.12 16.13 chr10 5556345 5557875 NM_005185CALML3 25.87 30.12 20.99 27.52 24.20 29.47 22.90 chr10 14856723 14857363NM_031453 FAM107B 7.73 10.25 7.01 7.39 10.11 6.09 5.50 chr10 2774239927743642 NM_001034842 PTCHD3 17.59 21.90 9.60 22.86 15.92 17.26 18.01chr10 37454095 37454949 NM_052997 ANKRD30A 12.21 11.95 9.38 13.30 14.1813.25 11.18 chr10 47216440 47217099 NM_001630 ANXA8L2 5.84 5.91 7.235.74 5.68 5.57 4.92 chr10 48447387 48447836 NM_015605 PTPN20B 6.11 4.807.59 6.05 6.40 6.48 5.34 chr10 94810041 94813282 NM_183374 CYP26C1 10.639.15 12.56 16.77 8.43 11.02 5.83 chr10 105207770 105209219 NM_001001412CALHM1 8.15 10.40 7.29 8.73 8.02 7.59 6.87 chr10 115928219 115929474NM_198795 TDRD1 17.76 22.47 16.00 22.82 18.87 16.13 10.26 chr10121285836 121286605 NM_002925 RGS10 5.73 6.90 5.00 5.45 4.86 5.35 6.79chr10 127453925 127455192 NM_147191 MMP21 8.25 8.96 7.79 9.38 9.73 6.547.08 chr10 127574241 127575495 NM_145235 FANK1 10.62 8.93 10.89 11.158.44 10.33 13.99 chr10 135086692 135087332 NM_001012508 SPRN 6.87 6.868.98 6.67 7.33 6.07 5.33 chr11 279861 280695 NM_025092 ATHL1 6.53 8.866.11 6.61 7.01 5.30 5.31 chr11 617645 618084 NM_021920 SCT 14.81 17.3910.06 14.50 13.85 16.29 16.80 chr11 820907 822857 NM_001039490 CD1516.38 5.06 6.13 7.94 5.60 7.86 5.70 chr11 1817998 1820162 NM_001145841TNNI2 7.95 5.26 9.55 8.00 8.60 9.31 7.01 chr11 1829198 1830654 NM_002339LSP1 9.03 10.85 10.43 8.71 10.86 8.30 5.02 chr11 1830796 1832555NM_002339 LSP1 8.89 10.18 9.69 7.43 12.21 7.07 6.78 chr11 18468081850966 NM_001013255 LSP1 11.19 10.57 9.06 12.27 15.93 12.00 7.33 chr112379793 2381347 NM_005706 TSSC4 7.76 9.51 8.09 6.60 9.69 7.70 5.00 chr112399707 2404341 NM_014555 TRPM5 5.66 5.89 6.14 6.36 5.71 5.29 4.56 chr112438393 2442742 NM_181798 KCNQ1 7.14 8.31 6.13 8.02 8.94 6.87 4.56 chr113195404 3196543 NR_027138 C11orf36 17.17 9.78 17.25 19.08 22.60 19.1815.13 chr11 3210151 3211310 NM_001039165 MRGPRE 20.44 29.55 17.71 25.5022.59 15.04 12.24 chr11 6217772 6218421 NM_001037329 CNGA4 20.21 26.5117.18 19.66 21.01 19.24 17.68 chr11 6297026 6299075 NM_145040 PRKCDBP8.97 11.95 7.50 10.60 9.31 9.25 5.23 chr11 17365816 17366370 NM_000525KCNJ11 8.13 8.84 8.27 9.94 9.19 5.28 7.26 chr11 60038866 60039824NM_001012417 MS4A13 15.15 20.20 17.78 9.47 18.28 14.50 10.69 chr1163829557 63830496 NM_004451 ESRRA 9.12 8.04 7.20 7.21 8.41 13.33 10.51chr11 66381905 66383149 NM_024036 LRFN4 9.46 10.98 8.44 10.91 9.16 10.087.17 chr11 76490756 76492103 NM_006189 OMP 16.14 25.55 13.79 17.01 16.5613.44 10.48 chr11 103412459 103413298 NM_001001711 DDI1 44.43 51.8239.03 48.69 45.01 39.40 42.65 chr11 111542612 111543474 NM_031275 TEX1213.56 17.44 9.35 15.76 15.43 10.55 12.85 chr11 119513243 119514010NM_012101 TRIM29 17.46 24.68 17.16 17.34 18.49 15.64 11.41 chr11128317836 128318785 NM_022112 TP53AIP1 6.44 7.93 6.88 7.28 5.92 5.555.10 chr12 56008 56752 NM_015232 IQSEC3 7.41 6.73 5.98 9.15 6.34 8.407.85 chr12 4424438 4425797 NM_020996 FGF6 18.12 25.21 14.09 20.15 18.4215.12 15.70 chr12 5023862 5024596 NM_002234 KCNA5 15.07 16.02 14.9017.75 14.33 18.36 9.05 chr12 5472728 5474382 NM_002527 NTF3 24.04 22.8223.82 23.24 23.26 27.27 23.84 chr12 31835936 31836485 NM_001013699LOC440093 31.58 32.60 28.81 33.36 33.59 27.22 33.90 chr12 3830584938306399 NM_001031748 C12orf40 6.41 8.52 5.37 5.21 6.56 4.90 7.88 chr1247462707 47463456 NM_015270 ADCY6 9.23 9.57 10.13 12.17 9.32 7.05 7.13chr12 50971013 50972263 NM_002281 KRT81 21.22 21.25 17.97 18.00 31.7621.71 16.61 chr12 50981226 50982470 NM_002284 KRT86 52.75 69.89 41.5385.40 52.68 30.08 36.94 chr12 51000856 51001605 NM_002282 KRT83 10.6317.74 7.45 10.68 10.07 9.89 7.94 chr12 51046887 51048336 NM_002283 KRT8533.42 41.59 26.55 33.64 40.47 30.66 27.61 chr12 79634253 79635392NM_005593 MYF5 7.31 6.69 7.12 7.51 6.78 6.20 9.53 chr12 8987288589873850 NM_152638 C12orf12 8.14 9.20 7.82 9.52 9.53 6.22 6.56 chr12100656881 100657620 NM_153694 SYCP3 19.04 18.20 18.83 13.89 23.00 21.4818.80 chr12 121780904 121781852 NM_032554 GPR81 10.91 13.15 8.07 13.2414.07 9.46 7.45 chr13 18653757 18654506 NM_006001 TUBA3C 24.48 29.3225.28 24.57 19.70 25.19 22.84 chr13 23780205 23781177 NM_178540 C1QTNF912.08 9.82 10.82 10.92 11.52 13.62 15.80 chr13 24235708 24236472NM_031277 RNF17 12.84 14.70 15.31 13.61 17.03 9.87 6.51 chr13 3568637835687238 NM_017826 SOHLH2 8.02 11.21 7.79 7.67 8.86 6.70 5.86 chr1336577620 36578169 NM_145203 CSNK1A1L 9.61 11.58 10.79 8.34 10.80 8.587.54 chr13 56612475 56613524 NM_001130407 LOC729250 42.01 47.96 29.6163.64 42.30 37.25 31.30 chr13 56618844 56620073 NM_001130407 LOC72925057.64 65.17 57.90 59.75 83.82 33.31 45.89 chr13 56625901 56626645NM_001130407 LOC729250 77.38 82.71 80.66 83.29 84.52 64.28 68.83 chr1356631990 56633239 NM_001130407 LOC729250 54.01 62.05 50.62 58.80 63.8057.68 31.09 chr13 56638553 56639782 NM_001130407 LOC729250 52.13 58.8649.60 66.82 61.00 43.23 33.29 chr13 94883460 94884537 NM_182848 CLDN108.83 7.22 9.61 8.47 11.83 7.52 8.33 chr13 97626570 97627629 NM_178861RNF113B 32.89 46.88 19.32 39.08 31.42 36.69 23.98 chr13 113155649113156614 NM_138430 ADPRHL1 13.03 14.48 20.29 9.81 11.31 9.94 12.37chr14 18622785 18623513 NR_027480 POTEG 31.88 30.01 40.36 37.79 31.5621.78 29.75 chr14 46190425 46191076 NM_080746 RPL10L 8.93 7.89 7.79 9.607.70 8.45 12.15 chr14 59263472 59264928 NM_206857 RTN1 26.09 33.37 19.9831.71 19.15 20.18 32.13 chr14 92882593 92883942 NM_182971 COX8C 30.3416.34 24.90 25.16 22.78 51.39 41.50 chr14 100262203 100262732 NM_003836DLK1 11.09 10.57 12.30 11.27 9.89 10.37 12.13 chr14 101097753 101098912NM_001362 DIO3 15.63 12.01 18.23 15.00 19.59 16.86 12.08 chr14 104266497104266926 NM_199165 ADSSL1 21.77 17.08 27.02 23.39 26.43 16.22 20.45chr15 19336475 19337416 NM_207355 POTEB 42.79 47.01 48.91 41.82 55.4137.77 25.81 chr15 20506397 20507346 NM_001033028 CYFIP1 20.89 27.0421.56 19.67 26.96 13.88 16.26 chr15 22471456 22472405 NM_018958 C15orf212.17 9.71 10.47 18.23 11.88 11.04 11.70 chr15 22750434 22752183NM_005678 SNURF 30.41 29.24 23.55 27.41 37.18 31.03 34.06 chr15 4172813541729101 NM_172097 CATSPER2 28.35 9.42 28.92 43.02 36.66 27.49 24.57chr15 57285847 57286806 NM_033195 LDHAL6B 14.17 13.16 14.91 14.06 12.4014.72 15.75 chr15 67009239 67010467 NM_145658 SPESP1 37.94 19.77 26.4346.95 43.55 45.96 44.97 chr15 71521869 71522920 NM_001042367 C15orf6013.97 14.42 8.82 14.37 17.14 12.57 16.48 chr15 72254477 72255116NM_201526 ISLR 20.56 26.49 22.67 21.15 26.10 14.24 12.72 chr16 675033675787 NM_001005920 JMJD8 12.92 14.80 10.24 14.96 13.55 11.61 12.34chr16 1068092 1070441 NM_001053 SSTR5 34.53 40.96 37.29 29.40 37.3638.88 23.29 chr16 1478290 1479239 NM_001013658 C16orf38 14.41 14.6615.85 13.68 17.47 14.01 10.79 chr16 1522860 1524624 NM_024600 TMEM2049.18 11.92 7.73 7.63 9.60 10.22 8.01 chr16 2807389 2808027 NM_144957PRSS21 10.34 9.77 6.95 15.92 13.20 10.41 5.77 chr16 3082682 3083229NM_032805 ZSCAN10 5.79 6.51 6.50 6.69 5.28 4.66 5.13 chr16 34330083434339 NM_001083601 NAT15 49.84 52.76 30.21 62.03 68.42 31.98 53.63chr16 3486239 3486788 NM_001080524 C16orf90 13.78 16.85 12.31 15.6714.16 11.34 12.34 chr16 32594155 32595099 NM_016212 TP53TG3 29.48 32.0219.45 39.59 30.74 26.26 28.82 chr16 33112997 33113931 NM_016212 TP53TG336.76 40.56 20.49 50.56 35.68 30.82 42.46 chr16 33169532 33170466NM_016212 TP53TG3 36.49 41.41 20.71 48.20 36.30 36.20 36.10 chr1645160325 45161269 NR_026556 ANKRD26P1 28.14 29.86 22.30 27.10 26.6737.82 25.07 chr16 54423975 54424824 NM_001266 CES1 13.58 18.06 10.2915.23 18.75 8.91 10.22 chr16 65156999 65158350 NM_052999 CMTM1 24.6931.78 23.76 23.95 21.11 21.31 26.23 chr16 65740729 65741468 NM_033309B3GNT9 9.80 5.64 11.37 11.74 8.35 11.84 9.86 chr16 66571411 66572180NM_001129758 DPEP3 13.68 14.14 11.24 16.10 15.37 13.19 12.03 chr1670080556 70081425 NM_006961 ZNF19 14.69 16.51 14.66 15.45 13.62 14.4513.44 chr16 73012597 73013956 NM_001011880 CLEC18B 6.59 7.74 6.68 7.057.54 5.79 4.76 chr16 73797516 73798755 NM_001025200 CTRB2 5.57 6.80 6.324.54 5.73 5.20 4.83 chr16 82781765 82782707 NM_001145400 ADAD2 14.1111.48 14.36 17.08 14.35 11.97 15.42 chr16 87330130 87330969 NM_001142864FAM38A 7.92 7.60 8.77 9.15 8.01 7.78 6.21 chr17 2645705 2646634NM_015085 GARNL4 14.28 17.59 17.08 10.76 12.46 18.52 9.25 chr17 33216393322576 NM_032598 SPATA22 51.95 55.52 34.18 58.36 57.16 55.68 50.81chr17 3446892 3447641 NM_018727 TRPV1 7.86 9.45 9.32 6.62 8.22 5.26 8.27chr17 4743347 4746194 NM_001145536 LOC100130311 12.04 11.16 13.76 15.3111.18 12.03 8.80 chr17 6839343 6841183 NM_000697 ALOX12 6.88 9.54 5.437.80 7.84 5.69 5.01 chr17 7697816 7699847 NM_203411 TMEM88 7.24 6.799.70 8.97 7.79 5.20 4.99 chr17 9748582 9750053 NM_002903 RCVRN 18.0619.85 20.69 13.99 18.70 15.67 19.45 chr17 17421328 17421768 NM_148173PEMT 7.16 8.10 7.27 6.57 8.21 5.86 6.93 chr17 18468941 18469890NM_182568 CCDC144B 22.52 22.23 20.82 21.07 25.97 26.48 18.55 chr1718795404 18796353 NM_152351 SLC5A10 10.73 13.89 11.21 10.17 10.18 10.598.35 chr17 20739556 20740720 NM_001004306 CCDC144NL 29.77 30.08 30.3129.12 36.76 29.55 22.82 chr17 21748717 21749646 NM_203392 FAM27L 15.1618.01 12.53 16.83 18.19 13.17 12.24 chr17 23658080 23658835 NM_173624FLJ40504 6.71 5.39 6.50 7.75 7.10 5.64 7.91 chr17 34250855 34251398NM_001080465 C17orf98 10.83 5.23 12.21 15.31 13.35 10.62 8.25 chr1736475474 36476433 NM_033184 KRTAP2-4 10.88 8.69 10.10 13.71 9.85 11.7011.20 chr17 36725274 36726223 NM_031964 KRTAP17-1 19.55 27.44 14.6322.33 24.07 15.44 13.42 chr17 36996489 36997433 NM_000526 KRT14 13.4815.74 13.74 13.64 11.94 14.69 11.14 chr17 37034156 37034805 NM_000422KRT17 6.67 7.86 5.51 5.86 7.48 7.43 5.89 chr17 37527672 37528921NM_033194 HSPB9 11.20 12.86 9.23 13.66 11.51 9.62 10.33 chr17 3919149939192250 NM_025237 SOST 7.00 7.08 6.97 6.63 7.00 8.97 5.34 chr1749254484 49255423 NM_032559 KIF2B 12.32 13.23 9.42 14.20 13.74 11.1612.19 chr17 62470387 62471721 NM_000727 CACNG1 22.03 25.91 22.92 18.2228.56 24.30 12.28 chr17 71815097 71816147 NM_032134 QRICH2 34.88 25.6531.36 32.78 39.30 42.92 37.29 chr17 75766293 75767047 NM_024110 CARD1411.13 11.97 11.32 8.22 16.82 7.37 11.09 chr17 75927527 75928476NM_020914 RNF213 8.67 10.87 6.06 10.11 7.92 11.12 5.93 chr17 7710974177110865 NM_012418 FSCN2 15.82 17.27 13.30 17.84 17.34 15.34 13.80 chr1777386013 77386662 NM_001007533 DYSFIP1 8.51 13.55 8.12 7.60 10.08 6.295.41 chr17 77473624 77474153 NM_032711 MAFG 11.26 7.11 8.03 16.55 10.9314.55 10.40 chr17 77925511 77926855 NM_018949 UTS2R 10.29 7.32 12.9010.94 12.78 10.75 7.07 chr18 569985 570939 NM_004066 CETN1 40.99 11.8845.66 55.56 50.67 39.81 42.34 chr18 13815665 13816754 NM_005913 MC5R24.82 31.04 25.88 21.22 35.13 17.21 18.47 chr18 18251681 18252650NM_172241 CTAGE1 17.99 19.71 16.34 24.18 24.73 8.77 14.19 chr18 2196731321968170 NM_001025096 PSMA8 17.28 19.59 17.74 16.87 17.36 14.62 17.52chr18 27594502 27595268 NM_001034172 MCART2 19.93 24.74 16.81 24.5321.00 12.87 19.65 chr18 42802973 42805327 NM_145653 TCEB3C 134.12 138.86129.94 145.26 146.36 120.71 123.57 chr18 42808886 42811242 NM_145653TCEB3C 130.36 139.99 125.33 143.84 141.45 130.20 101.36 chr18 4281502542816781 NM_016427 TCEB3B 44.78 43.66 36.25 44.78 43.21 52.83 47.94chr18 46062462 46063396 NM_015844 MBD1 11.75 21.26 8.42 11.86 8.59 8.9411.42 chr18 59294602 59295373 NM_002639 SERPINB5 7.41 8.82 5.33 8.856.76 7.45 7.23 chr18 75723859 75725028 NM_012283 KCNG2 23.25 23.66 19.8215.02 26.45 26.68 27.88 chr19 425784 426743 NM_182577 ODF3L2 8.57 9.137.41 8.54 13.27 6.26 6.78 chr19 1904572 1905206 NM_152771 C19orf34 10.8212.33 10.89 13.22 11.30 10.70 6.47 chr19 3129270 3130838 NM_003775 S1PR410.84 5.71 9.59 10.85 14.04 16.52 8.36 chr19 4743132 4744201 NM_018708FEM1A 18.37 21.90 13.30 17.30 20.23 23.20 14.30 chr19 5405666 5407310NM_181710 ZNRF4 24.45 25.36 27.26 24.38 22.44 25.60 21.66 chr19 57355225736971 NM_001134316 MGC24975 8.62 9.66 5.81 10.76 9.82 8.84 6.83 chr197652515 7653759 NM_001042461 TRAPPC5 14.36 8.56 15.54 12.75 17.37 23.498.48 chr19 8668814 8669778 NM_178525 ACTL9 15.26 23.38 9.71 14.47 15.2113.45 15.34 chr19 8794380 8795329 NM_144693 ZNF558 18.84 28.56 16.4014.72 22.21 13.46 17.68 chr19 10081535 10082279 NM_002566 P2RY11 22.2731.42 20.97 27.40 25.50 17.83 10.48 chr19 10082415 10083249 NM_002566P2RY11 12.04 16.24 10.64 15.02 12.36 11.88 6.13 chr19 15699554 15700503NM_013939 OR10H2 23.49 18.35 21.20 31.69 26.08 21.19 22.44 chr1915779233 15780184 NM_013940 OR10H1 29.97 40.31 29.73 34.01 27.70 29.1018.96 chr19 16861645 16862379 NM_003950 F2RL3 14.99 15.14 12.73 17.7819.96 15.40 8.95 chr19 17348683 17349427 NM_031310 PLVAP 12.02 14.1811.19 11.93 9.23 11.86 13.71 chr19 18406231 18409280 NM_016368 ISYNA19.47 11.10 12.24 9.39 7.89 8.91 7.27 chr19 19485956 19487705 NM_015965NDUFA13 23.44 19.43 23.04 29.97 24.90 27.36 15.96 chr19 3790225637902885 NM_001110822 TDRD12 10.24 15.30 7.33 9.64 10.82 12.05 6.31chr19 40533503 40535052 NM_005303 FFAR1 15.08 23.03 14.60 13.11 16.6113.06 10.09 chr19 40631857 40632626 NM_005306 FFAR2 6.71 8.06 6.73 5.847.21 5.50 6.94 chr19 43568464 43569908 NM_152657 GGN 11.85 12.47 12.9312.05 14.12 11.55 7.96 chr19 45587124 45588088 NM_144685 HIPK4 11.219.64 13.78 12.74 15.97 6.93 8.22 chr19 47040148 47041188 NM_001040283DMRTC2 16.20 18.08 16.69 15.23 16.11 11.39 19.71 chr19 51079188 51080617NM_015649 IRF2BP1 15.00 18.29 12.57 18.66 20.94 12.00 7.54 chr1951688686 51690541 NM_020709 PNMAL2 10.75 14.00 8.27 8.41 11.44 11.3611.01 chr19 51818645 51819269 NM_000960 PTGIR 8.01 9.63 6.81 9.42 8.557.21 6.46 chr19 53656820 53657569 NM_170720 KCNJ14 21.31 20.41 19.8423.04 20.77 28.49 15.28 chr19 54219222 54220176 NM_000737 CGB 16.1525.16 10.36 15.16 21.89 12.30 12.04 chr19 54226148 54226592 NM_033378CGB2 8.59 8.63 8.80 8.42 9.37 7.40 8.90 chr19 54238117 54239086NM_033043 CGB5 13.38 17.27 13.14 13.80 13.99 12.35 9.74 chr19 5424399154244945 NM_033183 CGB8 17.15 26.46 18.04 14.57 22.24 11.20 10.40 chr1954520006 54520660 NM_014037 SLC6A16 11.21 13.74 11.25 10.76 12.45 10.069.00 chr19 54583019 54583463 NM_144688 CCDC155 11.64 13.18 11.79 10.8511.00 13.09 9.92 chr19 55911639 55912709 NM_016148 SHANK1 8.56 11.197.77 10.46 8.58 8.24 5.11 chr19 56197491 56198350 NM_144505 KLK8 13.0213.57 12.16 6.85 24.82 10.45 10.28 chr19 58486057 58486682 NM_033341BIRC8 14.14 12.38 10.66 16.11 11.19 16.70 17.81 chr19 58732640 58733684NM_001079906 ZNF331 34.46 43.47 23.48 34.91 37.96 27.88 39.08 chr1958749250 58750079 NM_001079907 ZNF331 36.74 39.92 17.81 48.06 27.8640.94 45.89 chr19 60684392 60687751 NM_020378 NAT14 11.78 11.77 14.8610.43 10.57 13.11 9.93 chr19 61431236 61432273 NM_024303 ZSCAN5A 12.1810.85 13.96 15.95 13.91 9.26 9.17 chr19 62043095 62044373 NR_024059MIMT1 15.56 9.37 10.07 24.11 18.27 20.53 11.01 chr19 62321971 62322600NM_020903 USP29 14.56 19.05 7.49 16.71 9.10 20.35 14.65 chr19 6243351362434437 NM_001015878 AURKC 18.25 20.25 21.05 17.40 16.85 15.73 18.19chr2 10970124 10971578 NM_002236 KCNF1 27.57 35.95 25.50 22.03 35.3527.97 18.63 chr2 24250685 24252441 NM_001040710 C2orf84 8.45 7.67 6.8210.45 9.20 6.90 9.67 chr2 27226289 27226938 NM_175769 TCF23 15.75 15.0515.45 13.37 13.20 16.32 21.11 chr2 27351312 27351951 NM_173650 DNAJC5G9.14 10.47 10.01 6.46 10.06 11.69 6.17 chr2 48697663 48698722 NM_172196GTF2A1L 15.52 18.43 10.58 20.38 19.67 14.11 9.93 chr2 54336335 54337297NM_001003937 TSPYL6 16.41 22.39 20.92 15.91 15.12 9.41 14.72 chr284370521 84371670 NR_003663 LOC388965 39.46 51.89 36.84 42.03 36.9130.24 38.85 chr2 94900282 94901626 NM_144705 TEKT4 15.42 22.17 14.9413.43 18.51 14.64 8.84 chr2 96063947 96065201 NM_207328 LOC150763 55.7068.78 50.78 52.54 61.79 52.91 47.43 chr2 96145156 96146095 NM_000682ADRA2B 9.16 4.83 9.49 10.65 12.47 11.29 6.23 chr2 97716522 97717066NM_207519 ZAP70 13.84 13.90 18.74 12.51 13.22 12.61 12.03 chr2 9771722297718571 NM_207519 ZAP70 15.12 16.57 19.65 14.54 13.86 14.33 11.75 chr2105224921 105225570 NM_007227 GPR45 52.58 57.22 40.22 63.81 57.11 56.1040.99 chr2 108702582 108703121 NM_006267 RANBP2 13.69 14.48 11.27 13.7411.08 14.62 16.97 chr2 109909568 109910007 NM_005054 RGPD5 11.05 8.939.90 12.92 8.77 10.96 14.80 chr2 113972455 113974501 NM_012184 FOXD4L18.97 8.96 7.68 9.14 9.89 9.21 8.95 chr2 127679614 127680288 NM_001001665CYP27C1 7.19 7.01 5.90 7.70 8.54 6.68 7.33 chr2 130672072 130672916NM_207312 TUBA3E 32.99 51.31 51.63 35.18 7.97 6.77 45.08 chr2 131691617131692543 NM_001083538 POTEE 33.43 32.45 39.31 33.73 27.25 33.52 34.30chr2 131949675 131950614 NM_080386 TUBA3D 32.45 38.45 37.35 36.38 34.1826.17 22.18 chr2 132891446 132892097 NM_001508 GPR39 10.53 18.07 6.379.90 9.93 8.50 10.40 chr2 198358003 198359552 NM_197970 BOLL 17.79 12.0110.70 17.83 17.73 19.97 28.50 chr2 219961021 219961580 NM_012100 DNPEP7.33 6.58 8.34 6.30 6.46 7.28 9.02 chr2 232952384 232953053 NM_001632ALPP 18.16 18.22 16.24 18.67 14.64 22.65 18.53 chr2 232980448 232981097NM_031313 ALPPL2 21.79 25.63 20.09 18.23 23.31 29.65 13.82 chr2234316887 234318242 NM_001001394 DNAJB3 39.49 45.91 43.13 39.57 46.9035.98 25.44 chr2 238371764 238372800 NM_001080504 RBM44 28.42 7.37 28.4840.50 38.17 23.95 32.02 chr2 238713572 238714819 NM_198582 KLHL30 21.7530.96 20.71 21.12 25.44 14.63 17.65 chr2 241218250 241218894 NM_005301GPR35 15.19 16.38 16.96 17.77 14.29 15.30 10.43 chr20 4176544 4177393NM_000678 ADRA1D 10.90 6.25 12.43 12.00 11.32 14.71 8.69 chr20 2296373722964890 NM_001052 SSTR4 10.83 10.15 8.62 13.86 6.83 12.97 12.52 chr2023293995 23294644 NM_022482 GZF1 30.99 39.75 24.66 31.97 33.79 30.8224.93 chr20 23915334 23916273 NM_178312 GGTLC1 7.19 7.63 7.12 8.95 8.555.84 5.07 chr20 23917142 23917891 NM_178311 GGTLC1 18.89 26.49 16.6418.00 20.04 14.95 17.24 chr20 33343147 33344486 NM_178468 FAM83C 17.758.15 18.71 21.19 24.22 16.83 17.40 chr20 35582039 35583589 NM_005386NNAT 33.72 40.05 25.96 34.31 40.40 35.40 26.20 chr20 41575691 41577158NM_015478 L3MBTL 12.63 18.43 10.95 12.71 15.03 10.69 7.96 chr20 4241707942418028 NM_175914 HNF4A 7.53 10.97 6.28 7.09 7.82 6.95 6.07 chr2054541229 54541876 NM_001013646 C20orf107 6.65 7.80 6.67 5.37 5.94 6.497.63 chr20 54633696 54636839 NM_003222 TFAP2C 8.83 8.99 6.84 7.11 7.7215.10 7.24 chr20 55337555 55338610 NM_198265 SPO11 9.75 11.00 9.59 13.298.70 8.18 7.74 chr20 56847401 56850580 NM_016592 GNAS 31.50 38.68 26.0535.15 22.58 35.61 30.92 chr20 56859948 56860603 NM_080425 GNAS 11.5112.34 8.53 14.46 13.16 10.98 9.60 chr20 56860909 56861548 NM_080425 GNAS17.66 27.42 13.40 24.57 17.10 10.40 13.09 chr20 56861712 56864669NM_080425 GNAS 19.16 15.77 15.22 22.46 20.08 21.47 19.95 chr20 5719979957201264 NM_178457 ZNF831 11.74 6.05 10.06 16.51 16.27 11.77 9.78 chr2057941206 57941955 NM_014258 SYCP2 8.65 5.32 7.35 12.59 8.64 9.73 8.25chr20 61963314 61964663 NM_080622 C20orf135 12.67 9.75 10.65 11.76 20.1813.41 10.30 chr20 62207820 62209374 NM_005286 NPBWR2 26.26 24.42 22.4724.66 24.57 27.27 34.18 chr21 10120617 10121645 NM_001187 BAGE 7.9910.81 7.54 9.12 6.10 5.37 9.01 chr21 13903590 13904521 NM_174981 POTED51.34 46.21 57.76 45.85 60.93 54.30 42.99 chr21 13974048 13976108NM_001013729 LOC441956 77.69 73.54 92.14 65.11 66.59 89.88 78.90 chr2138414860 38415509 NR_026840 DSCR8 9.00 10.15 6.06 9.55 10.79 8.16 9.31chr21 42659517 42660181 NM_003225 TFF1 5.79 7.63 5.57 5.56 5.35 4.745.90 chr21 43902099 43903348 NM_015056 RRP1B 23.59 22.42 21.15 20.4124.32 22.09 31.16 chr21 44700420 44701794 NM_030891 LRRC3 18.18 21.4117.73 23.50 18.77 11.36 16.32 chr21 44784324 44785274 NM_198691KRTAP10-1 7.13 6.09 7.57 5.78 9.30 6.57 7.46 chr21 45700562 45701211NM_030582 COL18A1 11.83 10.72 15.92 10.55 12.80 9.86 11.14 chr2214667762 14668589 NM_001136213 POTEH 21.26 21.47 22.71 26.29 16.20 19.4121.44 chr22 15868297 15869926 NM_001037814 GAB4 11.96 15.42 10.68 10.1013.77 12.68 9.11 chr22 17499105 17499734 NM_053006 TSSK2 25.77 34.8125.99 21.26 33.82 17.68 21.03 chr22 18089100 18094254 NM_000407 GP1BB32.20 31.19 36.88 36.43 40.87 28.46 19.39 chr22 21317982 21318951NM_199127 GGTLC2 7.16 7.95 6.81 9.86 7.33 5.39 5.60 chr22 2446753924468284 NM_032608 MYO18B 10.70 14.80 12.67 13.04 10.09 6.91 6.67 chr2228033113 28034747 NM_152237 GAS2L1 7.52 9.44 7.26 7.05 7.12 7.69 6.59chr22 35231985 35232930 NM_001102371 FOXRED2 14.68 13.97 10.80 18.1418.55 14.52 12.14 chr22 40425282 40426136 NM_152513 MEI1 19.72 17.9318.81 22.21 28.01 19.62 11.75 chr22 43271073 43272028 NM_032287 LDOC1L10.60 10.75 11.41 10.09 11.50 10.33 9.49 chr22 45307998 45310932NM_014246 CELSR1 70.96 93.81 68.97 73.16 72.61 82.10 35.09 chr2245400538 45401974 NM_015124 GRAMD4 17.30 20.44 18.09 18.08 17.13 15.7714.26 chr22 48871561 48872205 NM_018995 MOV10L1 10.14 12.47 9.47 13.119.86 8.70 7.22 chr22 49099931 49100765 NM_001001794 FAM116B 5.91 5.655.29 6.21 5.47 6.88 5.94 chr3 10124574 10125513 NM_173472 C3orf24 12.8517.07 9.59 15.85 16.80 9.48 8.33 chr3 14835016 14835970 NM_152536 FGD57.71 8.35 6.89 7.29 10.79 5.16 7.77 chr3 16621582 16622837 NM_001351DAZL 21.35 16.87 15.93 32.41 18.72 22.63 21.52 chr3 42701821 42702885NM_152393 KBTBD5 8.83 9.26 5.65 10.79 12.31 8.79 6.19 chr3 4684990246851347 NM_182702 TESSP2 29.30 27.93 31.12 33.01 28.33 29.65 25.75 chr348673342 48674613 NM_001407 CELSR3 9.92 11.56 8.04 12.43 8.08 9.26 10.14chr3 49730772 49731516 NM_198722 AMIGO3 10.00 12.08 7.70 11.18 9.2513.20 6.59 chr3 49914941 49915590 NM_002447 MST1R 10.55 15.02 11.1710.62 11.91 8.19 6.38 chr3 110319501 110320450 NM_014429 MORC1 24.2326.10 24.02 27.27 26.25 19.43 22.30 chr3 120346987 120348436 NM_152539C3orf30 7.48 9.20 4.57 8.62 8.71 9.11 4.64 chr3 128189606 128191165NM_032242 PLXNA1 15.39 14.53 17.35 13.89 14.92 18.07 13.61 chr3129668583 129669442 NM_153330 DNAJB8 6.38 7.34 6.07 7.04 6.55 4.68 6.60chr3 139210899 139212158 NM_016369 CLDN18 18.57 23.04 14.67 22.74 21.7218.03 11.20 chr3 140221409 140222877 NM_001013650 LOC389151 41.01 28.3938.95 36.78 36.11 56.21 49.62 chr3 140245379 140247062 NM_001134657FLJ46210 37.02 18.35 28.21 44.87 32.74 53.71 44.25 chr3 142979153142980397 NM_139209 GRK7 23.34 28.43 20.20 29.03 26.26 21.24 14.90 chr3144163847 144164576 NM_198504 PAQR9 18.52 20.01 19.16 19.07 16.48 18.5917.82 chr3 185300280 185300841 NM_182589 HTR3E 12.08 15.07 9.43 16.8912.85 9.18 9.04 chr3 197549449 197550093 NM_138461 TM4SF19 9.74 8.8210.20 9.90 9.55 8.73 11.24 chr4 42930 43789 NM_001039127 ZNF718 39.9841.93 39.58 46.85 41.26 34.88 35.37 chr4 608783 610032 NM_000283 PDE6B10.38 14.95 8.37 10.13 12.11 9.33 7.41 chr4 688156 688920 NM_006315PCGF3 17.69 29.34 14.15 25.33 17.81 10.18 9.34 chr4 3219787 3221216NM_001012982 C4orf44 15.06 8.40 15.67 17.28 16.55 18.19 14.30 chr49392324 9394073 NM_000798 DRD5 42.67 43.41 34.80 47.18 43.39 43.82 43.39chr4 41678341 41679190 NM_001029955 WDR21B 40.08 40.97 41.45 42.09 35.3240.79 39.85 chr4 89837818 89838760 NM_153757 NAP1L5 16.90 37.22 11.4616.38 14.75 11.12 10.48 chr4 91978847 91979495 NM_183049 TMSL3 8.39 9.027.07 11.97 6.73 9.24 6.31 chr4 111616373 111617632 NM_001977 ENPEP 16.6917.61 20.70 14.01 19.59 15.78 12.45 chr4 123072986 123073715 NM_003305TRPC3 33.69 35.20 28.67 50.78 37.69 23.47 26.35 chr4 123519326 123520290NM_001159285 ADAD1 34.23 43.17 29.98 40.11 36.10 27.17 28.84 chr4128870244 128871472 NM_031291 SLC25A31 42.12 45.89 31.83 35.68 55.2445.25 38.81 chr4 151723471 151724530 NM_006439 MAB21L2 12.87 17.24 14.1013.78 11.54 13.09 7.45 chr4 155921173 155922811 NM_144979 RBM46 38.7528.10 28.66 52.28 40.85 43.39 39.23 chr4 164613968 164615243 NM_032136TKTL2 25.70 31.88 16.28 27.64 23.60 26.10 28.72 chr4 191098183 191099517NM_004477 FRG1 12.98 9.68 10.66 13.98 12.28 12.98 18.27 chr4 191142243191143786 NM_020040 TUBB4Q 24.43 27.81 24.37 31.81 26.05 19.51 17.06chr5 244817 245561 NM_001080478 LOC389257 5.97 5.16 6.12 5.96 6.56 6.815.22 chr5 1254190 1255119 NM_001003841 SLC6A19 7.07 7.45 6.99 6.48 8.268.15 5.08 chr5 1345979 1347540 NM_198255 TERT 16.69 18.46 18.32 18.3314.84 21.08 9.07 chr5 55069354 55070737 NM_001136034 DDX4 23.01 26.7517.21 23.30 22.58 25.53 22.70 chr5 101862110 101863456 NM_173488 SLCO6A18.79 8.43 8.06 9.00 13.43 6.48 7.31 chr5 121214963 121216013 NM_177478FTMT 19.38 12.09 17.31 23.23 22.13 17.32 24.18 chr5 135720291 135721827NM_020389 TRPC7 36.13 31.74 33.43 33.27 36.43 35.60 46.30 chr5 140032247140034004 NM_194249 DND1 12.88 13.55 16.66 12.73 17.44 9.92 6.99 chr5140663339 140663988 NM_031947 SLC25A2 6.85 6.80 6.72 7.52 7.55 6.87 5.63chr5 140699090 140699739 NM_018915 PCDHGA2 12.25 11.25 19.91 8.71 13.7213.53 6.36 chr5 140752377 140753116 NM_032088 PCDHGA8 6.51 5.93 5.057.79 6.55 8.22 5.52 chr5 140790699 140792828 NM_003735 PCDHGA12 19.4116.52 14.46 23.15 17.32 33.40 11.58 chr5 149525754 149527118 NM_001804CDX1 9.49 10.53 7.67 9.99 8.81 7.44 12.54 chr5 157010923 157012370NM_007017 SOX30 8.87 9.41 7.07 11.83 10.85 6.58 7.46 chr5 169464997169466263 NM_144769 FOXI1 8.68 9.08 8.77 8.37 11.06 9.80 5.00 chr5175041640 175042484 NM_022304 HRH2 5.93 6.93 5.74 7.88 4.49 4.39 6.15chr6 601409 602763 NM_148959 HUS1B 26.93 29.09 24.65 31.45 24.57 24.7927.02 chr6 3793831 3796095 NM_012135 FAM50B 28.32 48.91 10.94 39.4821.57 24.69 24.36 chr6 22676880 22678720 NM_138574 HDGFL1 52.66 73.9237.83 68.32 48.13 44.60 43.16 chr6 25834225 25835591 NM_170610 HIST1H2BA23.66 27.27 22.72 19.92 29.03 26.30 16.73 chr6 26354608 26355450NM_003547 HIST1H4G 23.76 27.28 23.68 26.93 28.01 16.90 19.77 chr629503310 29504241 NM_013937 OR11A1 6.79 7.98 7.66 7.89 5.89 6.91 4.41chr6 31799061 31799910 NM_138272 C6orf25 9.72 10.84 9.99 10.13 12.357.67 7.37 chr6 31891732 31893176 NM_005345 HSPA1A 13.53 19.66 9.92 12.5514.64 17.46 6.91 chr6 46763032 46764676 NM_001010870 TDRD6 19.97 10.4311.71 21.23 27.56 29.47 19.39 chr6 74120257 74121520 NM_001025290 DPPA518.02 19.56 25.53 18.75 12.81 15.57 15.91 chr6 74128640 74129269NM_001017361 C6orf221 9.75 12.28 10.70 9.06 11.83 7.00 7.64 chr674135930 74136609 NM_001080507 OOEP 15.83 18.38 12.19 15.05 15.64 18.6015.15 chr6 74161006 74161740 NM_018665 DDX43 19.23 20.89 17.30 19.0912.34 25.26 20.48 chr6 88813549 88814678 NM_030960 SPACA1 42.20 51.3918.60 45.44 33.13 49.52 55.11 chr6 112481472 112482226 NM_198239 WISP310.18 14.31 10.80 8.75 8.97 7.70 10.56 chr6 127838001 127839330NM_014702 KIAA0408 57.23 45.95 57.14 59.45 52.71 62.56 65.57 chr6160140502 160141661 NM_173516 PNLDC1 18.64 22.33 18.80 21.10 27.10 11.8310.67 chr6 160599193 160600752 NM_003058 SLC22A2 17.40 26.17 15.76 22.0113.69 14.66 12.10 chr7 1063268 1064522 NM_138445 GPR146 18.19 27.2521.97 15.76 18.52 12.01 13.65 chr7 2684888 2685852 NM_133463 AMZ1 9.6810.92 8.30 13.76 9.15 10.18 5.76 chr7 4135803 4136475 NM_001079653 SDK18.31 9.05 5.82 9.95 6.50 11.32 7.20 chr7 23715637 23716691 NM_032944STK31 36.46 44.10 32.08 48.20 36.80 31.60 25.99 chr7 53070155 53071394NM_182595 POM121L12 12.76 17.02 10.02 12.28 12.62 14.16 10.45 chr772077349 72078691 NM_198853 TRIM74 50.92 70.98 35.74 49.79 57.37 48.4343.20 chr7 72379763 72380692 NM_003602 FKBP6 53.21 58.89 50.39 53.8556.06 44.60 55.48 chr7 72486451 72488015 NM_003508 FZD9 13.83 14.5710.44 11.94 12.16 17.88 16.01 chr7 72882630 72883979 NM_001305 CLDN410.76 10.89 7.26 11.76 9.37 11.85 13.41 chr7 74862153 74863417 NM_198924TRIM73 64.84 79.30 54.71 64.21 67.97 57.16 65.69 chr7 116854422116855586 NR_023315 ASZ1 35.29 39.67 24.22 50.04 36.93 33.63 27.25 chr7129808524 129809253 NM_001868 CPA1 11.26 13.37 9.48 13.43 13.52 10.707.03 chr7 129917633 129920382 NM_002402 MEST 25.48 28.05 25.11 29.6327.20 21.73 21.17 chr7 142978433 142979168 NM_001008747 LOC441294 10.9512.33 9.95 12.17 9.57 9.22 12.48 chr7 149699704 149700939 NM_014374REPIN1 19.81 7.65 18.54 25.00 23.02 27.12 17.53 chr7 150376098 150377462NM_020322 ACCN3 14.81 20.46 11.26 15.42 19.76 12.07 9.86 chr7 154493038154494284 NM_024012 HTR5A 20.73 23.90 15.83 21.26 19.84 18.40 25.17 chr88597079 8598233 NM_194284 CLDN23 11.15 9.25 9.96 13.86 9.49 13.83 10.50chr8 21825506 21826155 NM_003974 DOK2 13.75 17.08 14.93 15.84 13.4811.21 9.95 chr8 22188020 22189349 NM_001135721 PIWIL2 26.18 32.45 22.3731.25 25.08 25.03 20.88 chr8 43265944 43266912 NM_001005365 POTEA 7.898.85 7.36 8.81 7.14 7.38 7.79 chr8 86760698 86762541 NR_003594 REXO1L2P74.86 97.36 61.15 85.63 98.07 48.47 58.51 chr8 88955074 88955655NM_152418 WDR21C 27.81 29.48 23.43 28.25 27.06 30.73 27.92 chr8143912441 143913671 NM_002066 GML 19.26 24.67 22.56 20.03 16.10 18.4613.75 chr8 145018352 145018996 NM_031308 EPPK1 9.82 12.17 9.24 10.3611.31 9.47 6.40 chr9 106804 109189 NM_207305 FOXD4 9.02 12.94 8.09 10.016.60 8.38 8.11 chr9 2707903 2708962 NM_133497 KCNV2 27.14 37.08 11.7328.00 18.16 40.68 27.22 chr9 5175227 5176457 NM_007179 INSL6 51.27 72.5056.04 51.63 51.21 31.22 44.99 chr9 42357193 42358439 NM_001012419ANKRD20A3 7.60 5.98 8.23 9.35 7.56 7.79 6.71 chr9 42708104 42709237NM_199244 FOXD4L4 7.11 7.48 5.99 7.06 7.49 8.13 6.52 chr9 4312372743124986 NM_001012421 ANKRD20A2 7.62 8.32 8.41 8.30 5.86 6.92 7.88 chr967515302 67516141 NM_001012419 ANKRD20A3 9.00 7.86 10.22 8.70 9.62 7.629.96 chr9 68490213 68491448 NM_001085476 FOXD4L6 10.18 10.48 7.32 11.7810.65 12.88 7.99 chr9 69667338 69668675 NM_199244 FOXD4L4 9.35 8.75 7.2812.34 9.31 10.91 7.53 chr9 70106903 70108958 NM_199135 FOXD4L3 9.2610.48 5.83 9.24 9.84 10.67 9.48 chr9 70818133 70818792 NM_002732 PRKACG13.34 12.92 12.08 17.39 12.64 14.12 10.91 chr9 103396630 103397877NM_147180 PPP3R2 18.30 23.07 19.92 21.89 14.86 16.26 13.82 chr9110657158 110658802 NM_006686 ACTL7B 47.75 59.69 42.00 50.58 52.28 46.3035.65 chr9 135433836 135434510 NM_001080515 FAM163B 10.11 14.29 13.6811.11 7.08 7.77 6.69 chr9 135490511 135491455 NM_000787 DBH 7.64 10.606.42 8.26 10.22 5.59 4.77 chr9 137552429 137553278 NM_002297 LCN1 6.116.44 5.54 6.63 7.75 5.01 5.30 chr9 138858099 138861468 NM_001080482LOC389813 15.49 10.55 16.63 23.08 10.98 13.18 18.53 chrX 169244 169903NM_012227 GTPBP6 17.18 23.22 9.47 16.37 17.65 21.36 15.03 chrX 170609171148 NM_012227 GTPBP6 11.18 12.30 10.26 10.94 12.61 12.21 8.77 chrX1673869 1674498 NM_004043 ASMT 18.69 22.83 15.76 22.90 18.02 13.94 18.67chrX 18148158 18149002 NM_153346 BEND2 54.90 45.78 46.81 63.36 58.0879.46 35.89 chrX 18149156 18149705 NM_153346 BEND2 16.82 14.17 15.1222.50 20.73 17.09 11.31 chrX 21583950 21587200 NM_153270 KLHL34 17.1916.12 13.40 16.65 11.57 31.45 13.95 chrX 21783926 21784775 NM_206923 YY222.96 26.31 9.09 26.27 21.62 29.42 25.03 chrX 27735245 27736219NM_182506 MAGEB10 10.81 16.08 8.36 9.57 11.64 9.40 9.83 chrX 3017441130175660 NM_177404 MAGEB1 25.68 34.17 16.93 27.11 29.73 22.66 23.45 chrX30999683 31000252 NM_031894 FTHL17 11.27 10.99 10.29 17.15 13.18 8.387.64 chrX 34058424 34059693 NM_203408 FAM47A 68.87 86.96 30.50 86.5035.83 91.33 82.10 chrX 36936001 36936845 NM_001013736 FAM47C 20.67 23.1018.45 19.48 20.62 18.42 23.98 chrX 44587800 44588965 NM_022076 DUSP2118.92 15.84 15.23 18.97 23.29 18.38 21.83 chrX 48050422 48051291NM_174962 SSX9 6.13 7.27 5.87 6.30 6.46 5.22 5.66 chrX 48156111 48156755NM_001034832 SSX4B 8.29 8.80 6.72 8.19 11.05 6.49 8.47 chrX 4834183448342497 NM_017883 WDR13 18.28 21.49 7.06 10.95 13.87 45.48 10.85 chrX51165707 51167656 NM_203407 CXorf67 30.06 26.64 29.35 29.25 30.28 29.6535.19 chrX 52255448 52255907 NM_001097603 XAGE1E 15.51 18.30 15.91 13.6413.99 16.40 14.80 chrX 52276731 52277175 NM_020411 XAGE1D 16.11 17.9815.41 18.66 16.36 14.24 13.99 chrX 52528399 52528858 NM_001097603 XAGE1E15.58 18.77 16.59 16.27 12.57 14.89 14.38 chrX 52549671 52550115NM_020411 XAGE1D 17.06 16.87 17.12 19.40 16.96 14.91 17.10 chrX 5256256552563009 NM_020411 XAGE1D 16.04 17.61 16.98 16.91 17.95 14.70 12.09 chrX52752781 52753645 NM_003147 SSX2 11.21 11.18 10.03 9.54 14.42 10.7511.35 chrX 52796355 52796804 NM_003147 SSX2 8.08 7.33 6.05 9.61 10.117.78 7.60 chrX 52857157 52858012 NM_130775 XAGE5 27.36 36.00 25.93 30.2329.31 16.74 25.93 chrX 52912981 52914510 NM_133179 XAGE3 17.89 20.8816.78 16.36 18.48 17.19 17.62 chrX 55117439 55118384 NM_001015038 PAGE2B8.81 10.48 9.18 7.68 9.73 6.09 9.70 chrX 55131445 55132374 NM_207339PAGE2 9.52 9.86 9.62 11.23 12.40 5.98 8.04 chrX 55263129 55263764NM_001013435 PAGE5 10.17 12.20 7.56 12.50 12.84 7.98 7.94 chrX 6919848969200038 NM_207320 OTUD6A 33.00 33.09 27.30 34.12 34.16 41.48 27.87 chrX73440474 73441108 NM_203303 ZCCHC13 12.81 13.74 10.46 11.17 16.43 15.329.71 chrX 89063104 89063753 NM_138960 TGIF2LX 10.59 12.27 7.49 11.6110.35 6.96 14.84 chrX 102078480 102079429 NM_001031834 RAB40AL 8.30 9.495.80 8.41 8.73 8.19 9.18 chrX 103154408 103155677 NM_001002916 H2BFWT24.83 22.71 21.06 23.44 22.93 35.42 23.40 chrX 119094651 119095297NM_032498 RHOXF2 20.02 23.85 14.66 20.84 13.48 29.42 17.88 chrX119176907 119177556 NM_001099685 RHOXF2B 17.65 22.01 11.11 23.40 11.9025.91 11.54 chrX 125513403 125515157 NM_178470 WDR40B 10.39 9.37 6.6710.16 6.79 24.20 5.12 chrX 132179867 132180816 NM_016521 TFDP3 26.4638.56 12.65 40.28 31.43 25.11 10.75 chrX 134802666 134803605 NM_018666SAGE1 9.00 11.98 8.52 10.38 8.94 6.35 7.82 chrX 139923943 139924587NM_139019 SPANXF1 6.42 7.29 5.56 6.09 6.06 6.11 7.38 chrX 140164130140164970 NM_022661 SPANXC 8.14 7.39 6.91 8.42 8.46 7.76 9.90 chrX140500305 140500869 NM_013453 SPANXA1 8.44 10.10 7.36 6.11 6.70 9.2611.09 chrX 140504824 140505753 NM_013453 SPANXA1 8.60 9.24 8.46 7.509.77 7.10 9.52 chrX 140614068 140614937 NM_145665 SPANXE 8.98 7.94 7.8410.97 9.14 8.10 9.86 chrX 146869951 146870810 NM_152578 FMR1NB 8.8510.34 6.35 7.08 7.03 11.36 10.93 chrX 152592892 152593731 NM_001039582PNCK 7.58 9.74 5.68 6.25 10.39 9.25 4.20 chrX 153062148 153063092NM_020061 OPN1LW 14.39 21.95 18.35 12.02 13.99 11.20 8.82 chrX 153176437153177581 NM_012253 TKTL1 21.48 31.34 16.87 21.52 19.53 28.13 11.51 chrX153465818 153466862 NM_139250 CTAG1A 14.38 11.35 11.43 10.83 14.24 29.089.38 chrX 153500530 153501564 NM_001327 CTAG1B 12.64 10.65 11.44 9.7313.54 19.87 10.63 chrX 153534848 153535884 NM_020994 CTAG2 16.20 15.4113.14 19.12 14.37 24.40 10.78 chrX 153765716 153766871 NM_001007524 F8A38.49 10.35 6.27 7.00 7.13 12.69 7.47 chrX 154262827 154263982NM_001007524 F8A3 9.26 12.96 7.00 10.82 6.47 12.48 5.87 chrX 154342433154343563 NM_001017991 H2AFB2 7.74 8.83 5.64 7.68 6.44 10.98 6.84 chrX154763420 154764572 NM_001145149 VAMP7 10.52 10.43 10.18 15.57 10.278.47 8.18 chrY 169244 169903 NM_012227 GTPBP6 18.35 24.35 11.54 15.5818.73 26.38 13.52 chrY 170609 171148 NM_012227 GTPBP6 11.32 12.22 8.1110.13 13.55 12.21 11.73 chrY 1673869 1674498 NM_004043 ASMT 20.62 26.6216.35 21.79 19.50 16.29 23.19 chrY 14607596 14608245 NM_004679 VCY 7.267.00 5.61 7.58 6.21 7.53 9.64 chrY 14676912 14677741 NM_181880 VCY1B16.85 16.64 13.82 16.24 16.71 18.58 19.09 chrY 57622620 57623772NM_001145149 VAMP7 10.81 10.80 14.79 14.35 8.30 8.42 8.18

TABLE 5 Frequently methylated 5′ gene ends in brain tumors. PeaksAverage P No. of Tumors Downstream Gene Within chr Start End value scorewith Peaks Strand Start End Symbol Strand Start chr1 38002438 3800358710.10 12 chr1 60311689 60312524 9.01 16 chr1 63556597 63560345 8.49 15chr1 150078663 150079332 11.19 11 + 150077568 chr1 153557083 1535578278.77 11 − 153556874 153560562 C1orf104 chr1 179719350 179719904 10.69 13chr1 181421423 181422187 9.18 19 chr1 207914587 207915953 6.93 9 +207915292 207916358 G0S2 + 207915292 chr1 222870205 222872650 9.25 10 +222870801 chr1 234752777 234754328 9.67 11 + 234748187 chr10 4641247746413703 7.66 21 chr10 50486546 50487915 9.95 11 chr10 50488946 5049171510.13 15 + 50488352 50490772 SLC18A3 + 50488352 chr10 79066119 790667739.27 12 − 78314641 chr10 120344105 120345734 7.81 11 − 120342905 chr10124628733 124629607 7.80 12 chr11 10428000 10428654 7.76 16 chr1131789500 31795000 8.20 10 chr11 32411453 32412722 13.77 15 − 32365900chr11 63823533 63825290 17.92 13 + 63815368 63824079 KCNK4 chr1242231009 42232738 12.35 17 chr12 47975997 47977646 11.74 21 + 47975175chr12 55168271 55169242 10.27 19 chr12 55918204 55919848 21.03 9 −55914952 chr12 79626169 79626805 12.87 20 + 79625576 79627382 MYF6 +79625576 chr12 84197229 84198765 8.80 11 chr13 40928925 40929564 6.50 17chr13 52211145 52212607 7.69 14 chr13 102224101 102224965 10.63 16 chr1436122209 36123483 8.76 9 chr14 36196539 36197988 8.90 13 + 36196532chr14 53491636 53492707 12.27 14 − 53486204 chr14 56344682 56346630 8.369 − 56337177 chr14 60021741 60022400 6.81 12 chr14 60045099 600479429.30 11 + 60045690 60048278 SIX6 + 60045690 chr14 73776450 73777601 6.388 + 73775927 chr14 74147083 74147922 9.41 19 − 74034638 chr14 9430439094305319 7.99 17 − 94304312 94306252 GSC − 94304312 chr14 105023673105024599 8.21 12 chr15 50867775 50868924 6.92 18 − 50836644 chr1572207930 72209773 11.51 13 + 72208767 chr15 81112195 81112934 8.26 16 −81009005 chr15 82113943 82114802 6.20 20 chr16 62621 63380 5.69 14 chr1619804086 19804635 8.37 20 chr16 31014172 31014616 5.57 13 chr16 5352334253524273 7.02 16 + 53522611 chr16 55229290 55229739 8.87 19 chr1655234406 55235250 6.28 12 chr17 4635389 4636518 8.20 10 − 46353204636469 VMO1 − 4635320 chr17 4638694 4639358 7.56 13 chr17 68666276868275 12.02 20 + 6867092 chr17 6886114 6887363 7.45 14 − 68856726887966 SLC16A11 − 6885672 chr17 36937436 36938490 8.11 8 chr17 4132993141330876 7.71 22 + 41328944 chr17 42251317 42252051 10.35 11 chr1744564528 44565791 10.73 10 chr17 55853774 55854910 6.09 16 chr1769719958 69720487 9.44 11 − 69717735 chr17 69781509 69782358 7.08 11chr17 77479685 77480419 7.24 13 + 77478995 chr18 5186243 5187292 11.7712 − 5133671 chr18 26875494 26877443 12.24 16 − 26824049 chr18 5325456553259819 10.42 18 + 53253914 chr18 55091928 55092557 11.94 15 chr1872664418 72665346 7.60 10 chr19 1812475 1814434 8.50 13 − 1803398 chr199469724 9470804 10.30 11 chr19 10258672 10259721 8.63 13 + 1025864910260198 ICAM4 + 10258649 chr19 10558593 10559142 6.31 12 chr19 1838986018391505 11.85 15 chr19 40222452 40224104 9.89 21 + 40213373 40223193SCN1B chr19 49455836 49456370 7.46 11 chr19 53891560 53892099 7.52 10 +53891039 chr19 59157320 59157774 9.54 19 chr2 10100088 10101856 10.60 16chr2 72226653 72228187 10.04 12 − 72209874 chr2 74582914 74584780 8.71 8− 74585677 74588329 PCGF1 + 74583251 chr2 111591681 111594348 13.30 21 +111206620 111592270 ACOXL chr2 115636321 115637260 13.24 12 + 115636201chr2 119322147 119323502 10.24 13 chr2 119630604 119631968 10.75 17 −119630288 119632941 C1QL2 − 119630288 chr2 119905436 119906957 8.58 13 +119905915 chr2 155263352 155264303 12.69 17 chr2 158161857 1581625097.18 12 − 158091524 chr2 160626827 160628085 6.25 14 chr2 172673432172675478 7.47 13 − 172672411 chr2 176653956 176656685 9.31 11 −176653080 176656936 EVX2 − 176653080 chr2 176671999 176672948 7.88 16 +176672775 176673734 HOXD12 + 176672775 chr2 176690151 176690900 10.3117 + 176689737 chr2 176737592 176738247 9.42 16 + 176737050 chr2200041833 200043073 13.73 23 + 200041065 chr2 222869621 222872193 7.9714 + 222871109 chr2 232103046 232104293 8.25 15 chr2 232279406 23228074513.71 9 chr2 233442461 233443405 8.93 17 chr20 22505521 22506890 10.2314 − 22489193 chr20 25076779 25077618 6.77 13 chr20 41251999 4125276310.05 12 chr20 41976531 41978482 10.08 20 + 41976905 chr20 4346953843470502 10.23 12 + 43468650 chr20 43531029 43532764 7.57 11 chr2121291294 21291968 10.35 10 chr21 34962773 34964708 7.05 10 + 34963557chr22 28039290 28041531 13.41 21 − 28038921 28041748 RASL10A − 28038921chr22 36807805 36808832 5.62 12 − 36804089 chr3 2115949 2117089 11.08 19chr3 32254374 32255424 7.95 11 chr3 50357760 50358604 8.66 9 chr3138020672 138021936 18.36 10 + 138020550 chr3 155629021 155629670 10.3510 − 155538154 chr3 159303324 159304476 14.83 12 − 159296493 chr3161649899 161651150 6.32 16 chr3 173648073 173649732 9.01 18 chr3188870393 188870937 13.49 19 chr4 5945893 5946532 7.96 15 chr4 5261217352613018 7.73 17 chr4 54661877 54662826 14.11 17 + 54661004 54662879GSX2 + 54661004 chr4 85636686 85637940 7.21 15 − 85633459 chr4 122211474122212323 18.69 9 − 122176231 chr4 174688523 174689572 9.32 15 +174688183 chr5 2805770 2806724 9.19 18 + 2805261 chr5 32747979 3275022912.42 16 + 32747421 chr5 37872516 37875378 15.39 13 − 37851509 chr563292326 63293075 10.85 20 chr5 76151227 76151876 8.32 17 + 76150588chr5 76968098 76969322 7.73 9 − 76960292 chr5 140286488 140287347 15.6217 + 140242037 chr5 140742560 140743119 8.07 13 + 140733834 chr5140747458 140748016 8.51 17 + 140742650 chr5 140767256 140768329 11.3619 + 140762703 chr5 145698688 145700318 10.46 12 + 145698779 145700276POU4F3 + 145698779 chr5 157030866 157031611 9.76 19 chr5 158690162158691536 7.47 12 chr6 10994972 10995711 7.30 13 chr6 26125798 261267589.25 13 chr6 27948718 27949880 10.00 11 − 27948904 27949268 HIST1H4L −27948904 chr6 30050074 30050923 5.05 13 chr6 30818753 30820599 10.08 16− 30818954 30820306 IER3 − 30818954 chr6 43720000 43721360 8.21 12 chr678228956 78230415 10.96 17 chr6 83130068 83131208 8.72 14 chr6 112147783112148610 9.95 13 − 112089177 chr6 117192435 117193290 8.00 14 −117180053 chr6 137857127 137857980 13.22 14 chr6 150326335 15032818432.01 19 + 150326835 chr7 526958 527907 9.01 15 chr7 5077779 50787276.16 15 + 5078263 5079380 LOC389458 + 5078263 chr7 27100570 271023008.47 12 − 27099138 chr7 27156749 27161918 7.67 13 − 27159862 27162821HOXA7 − 27159862 chr7 27185469 27186510 12.70 12 − 27187300 27191360HOXA11 chr7 27190648 27192605 14.02 16 + 27191551 chr7 27248657 272532188.83 9 + 27248688 27252717 EVX1 + 27248688 chr7 51352205 51352779 7.0820 chr7 116906858 116907422 10.33 12 chr7 120755525 120756474 10.97 17 +120752656 chr7 129912506 129914030 8.21 18 − 129914133 129918249 MESTIT1chr7 131983416 131984045 7.46 10 chr7 156125196 156126661 8.12 9 chr810626015 10628284 7.94 10 chr8 23618422 23620876 8.71 14 − 23615909 chr824826911 24828465 8.26 19 + 24827178 chr8 24868882 24870630 9.36 13 −24864387 chr8 26777575 26780521 15.03 22 − 26683138 chr8 5752221457523567 8.06 16 chr8 65873955 65874718 9.27 15 chr8 71144440 711471786.16 11 − 71126576 chr8 98359200 98360130 6.68 12 chr8 102573379102574213 9.31 19 chr8 110725080 110726354 9.55 13 − 110655580 chr934369432 34371281 7.87 20 − 34369016 34387849 C9orf24 − 34369016 chr985760867 85761816 9.92 8 chr9 90339041 90340305 7.18 17 chrX 111211507111212571 11.31 13 − 110904197 Peaks Gene Upstream Gene chr Start EndEnd Symbol Strand Start End Symbol chr1 38002438 38003587 − 3799853138003411 EPHA10 chr1 60311689 60312524 − 60228653 60312014 C1orf87 chr163556597 63560345 + 63561317 63563385 FOXD3 chr1 150078663 150079332150083264 LOC100132111 − 150076962 150079657 LOC100191040 chr1 153557083153557827 + 153557263 153567533 RUSC1 chr1 179719350 179719904 +179719338 180037338 CACNA1E chr1 181421423 181422187 + 181421796181480885 LAMC2 chr1 207914587 207915953 207916358 G0S2 + 207915292207916358 G0S2 chr1 222870205 222872650 222994872 CNIH3 + 222870801222994872 CNIH3 chr1 234752777 234754328 234779619 LGALS8 + 234753699234779619 LGALS8 chr10 46412477 46413703 + 46413551 46420574 GPRIN2chr10 50486546 50487915 + 50488352 50490772 SLC18A3 chr10 5048894650491715 50490772 SLC18A3 + 50492088 50543156 CHAT chr10 7906611979066773 79067583 KCNMA1 − 78314641 79067583 KCNMA1 chr10 120344105120345734 120345150 PRLHR − 120342905 120345150 PRLHR chr10 124628733124629607 + 124629138 124648220 LOC399815 chr11 10428000 10428654 +10428799 10485702 AMPD3 chr11 31789500 31795000 − 31768059 31789361 PAX6chr11 32411453 32412722 32413663 WT1 − 32365900 32413663 WT1 chr1163823533 63825290 + 63824438 63828815 C11orf20 chr12 42231009 42232738 −42034278 42231991 ADAMTS20 chr12 47975997 47977646 47978748 PRPH +47975175 47978748 PRPH chr12 55168271 55169242 − 55151002 55168448 GLS2chr12 55918204 55919848 55920742 NDUFA4L2 − 55914952 55920742 NDUFA4L2chr12 79626169 79626805 79627382 MYF6 + 79625576 79627382 MYF6 chr1284197229 84198765 + 84198166 84219692 ALX1 chr13 40928925 40929564 +40929541 40943013 C13orf15 chr13 52211145 52212607 − 52175399 52211948LECT1 chr13 102224101 102224965 − 102216465 102224150 C13orf27 chr1436122209 36123483 − 36118966 36121537 NKX2-8 chr14 36196539 3619798836216762 PAX9 + 36196532 36216762 PAX9 chr14 53491636 53492707 53493304BMP4 − 53486204 53493304 BMP4 chr14 56344682 56346630 56346937 OTX2 −56337177 56346937 OTX2 chr14 60021741 60022400 − 59972426 60022517C14orf39 chr14 60045099 60047942 60048278 SIX6 + 60045690 60048278 SIX6chr14 73776450 73777601 73799194 VSX2 + 73775927 73799194 VSX2 chr1474147083 74147922 74148787 LTBP2 − 74034638 74148787 LTBP2 chr1494304390 94305319 94306252 GSC − 94304312 94306252 GSC chr14 105023673105024599 + 105024301 105026169 CRIP1 chr15 50867775 50868924 50869501ONECUT1 − 50836644 50869501 ONECUT1 chr15 72207930 72209773 72216196ISLR2 + 72209795 72216196 ISLR2 chr15 81112195 81112934 81113783 CPEB1 −81009005 81113783 CPEB1 chr15 82113943 82114802 + 82113841 82499597ADAMTSL3 chr16 62621 63380 − 48057 62629 RHBDF1 chr16 19804086 19804635− 19777793 19803652 GPRC5B chr16 31014172 31014616 − 31009675 31013777VKORC1 chr16 53523342 53524273 53525896 IRX5 + 53522611 53525896 IRX5chr16 55229290 55229739 + 55230078 55231500 MT1A chr16 5523440655235250 + 55235216 55236353 MT1DP chr17 4635389 4636518 4636469 VMO1 −4635320 4636469 VMO1 chr17 4638694 4639358 + 4638993 4640425 GLTPD2chr17 6866627 6868275 6873685 BCL6B + 6867092 6873685 BCL6B chr176886114 6887363 6887966 SLC16A11 − 6885672 6887966 SLC16A11 chr1736937436 36938490 − 36933394 36938167 KRT19 chr17 41329931 4133087641331959 LOC100130148 + 41328944 41331959 LOC100130148 chr17 4225131742252051 − 42196856 42251081 WNT3 chr17 44564528 44565791 + 4456532744602350 B4GALNT2 chr17 55853774 55854910 + 55854646 55863568 C17orf64chr17 69719958 69720487 69721055 MGC16275 + 69721290 69769752 TTYH2chr17 69781509 69782358 + 69781991 69822617 DNAI2 chr17 7747968577480419 77481919 LOC92659 + 77478995 77481919 LOC92659 chr18 51862435187292 5187255 LOC642597 − 5133671 5187255 LOC642597 chr18 2687549426877443 26876779 DSC3 − 26824049 26876779 DSC3 chr18 53254565 5325981953309528 ONECUT2 + 53253914 53309528 ONECUT2 chr18 55091928 55092557 −55085246 55091605 RAX chr18 72664418 72665346 + 72665103 72811670 ZNF236chr19 1812475 1814434 1814564 KLF16 − 1803398 1814564 KLF16 chr199469724 9470804 − 9438030 9470279 ZNF560 chr19 10258672 1025972110260198 ICAM4 + 10258649 10260198 ICAM4 chr19 10558593 10559142 −10544347 10558991 AP1M2 chr19 18389860 18391505 + 18391135 18406459SSBP4 chr19 40222452 40224104 + 40223249 40249315 HPN chr19 4945583649456370 + 49455915 49471308 ZNF233 chr19 53891560 53892099 53901003FUT2 + 53891039 53901003 FUT2 chr19 59157320 59157774 + 5915810559177951 CACNG8 chr2 10100088 10101856 + 10101132 10112414 KLF11 chr272226653 72228187 72228471 CYP26B1 − 72209874 72228471 CYP26B1 chr274582914 74584780 74585700 LOC151534 + 74583251 74585700 LOC151534 chr2111591681 111594348 + 111594961 111642493 BCL2L11 chr2 115636321115637260 116318406 DPP10 + 115636201 116318406 DPP10 chr2 119322147119323502 − 119316216 119322229 EN1 chr2 119630604 119631968 119632941C1QL2 − 119630288 119632941 C1QL2 chr2 119905436 119906957 119912566TMEM37 + 119905915 119912566 TMEM37 chr2 155263352 155264303 + 155263338155421260 KCNJ3 chr2 158161857 158162509 158193645 ACVR1C − 158091524158162318 ACVR1C chr2 160626827 160628085 − 160510571 160627367 PLA2R1chr2 172673432 172675478 172675724 DLX2 − 172672411 172675724 DLX2 chr2176653956 176656685 176656936 EVX2 − 176653080 176656936 EVX2 chr2176671999 176672948 176673734 HOXD12 + 176672775 176673734 HOXD12 chr2176690151 176690900 176692916 HOXD10 + 176689737 176692916 HOXD10 chr2176737592 176738247 176746072 HOXD3 + 176737050 176746072 HOXD3 chr2200041833 200043073 200045724 FLJ32063 + 200041065 200045724 FLJ32063chr2 222869621 222872193 222878180 CCDC140 + 222871109 222878180 CCDC140chr2 232103046 232104293 − 232096114 232103426 NMUR1 chr2 232279406232280745 + 232281478 232286494 PTMA chr2 233442461 233443405 +233443237 233449351 UNQ830 chr20 22505521 22506890 22507280 C20orf56 −22489193 22507280 C20orf56 chr20 25076779 25077618 − 25069433 25077426LOC284798 chr20 41251999 41252763 − 40134805 41251971 PTPRT chr2041976531 41978482 42131668 TOX2 + 41978195 42131668 TOX2 chr20 4346953843470502 43472661 DBNDD2 + 43470328 43472661 DBNDD2 chr20 4353102943532764 + 43531807 43543586 WFDC2 chr21 21291294 21291968 + 2129250321833085 NCAM2 chr21 34962773 34964708 35012389 CLIC6 + 3496355735012389 CLIC6 chr22 28039290 28041531 28041748 RASL10A − 2803892128041748 RASL10A chr22 36807805 36808832 36809116 SLC16A8 − 3680408936809116 SLC16A8 chr3 2115949 2117089 + 2117246 3074645 CNTN4 chr332254374 32255424 + 32255174 32386817 CMTM8 chr3 50357760 50358604 −50353540 50358160 ZMYND10 chr3 138020672 138021936 138057424 TMEM22 +138020550 138057424 TMEM22 chr3 155629021 155629670 155630198 GPR149 −155538154 155630198 GPR149 chr3 159303324 159304476 159306630 SHOX2 −159296493 159303520 SHOX2 chr3 161649899 161651150 − 161635984 161650320TRIM59 chr3 173648073 173649732 − 173648027 173648897 GHSR chr3188870393 188870937 − 188869387 188870895 SST chr4 5945893 5946532 −5873391 5945686 CRMP1 chr4 52612173 52613018 + 52612349 52658215 SPATA18chr4 54661877 54662826 54662879 GSX2 + 54661004 54662879 GSX2 chr485636686 85637940 85638411 NKX6-1 − 85633459 85638411 NKX6-1 chr4122211474 122212323 122213123 C4orf31 − 122176231 122213123 C4orf31 chr4174688523 174689572 174699556 NBLA00301 + 174688183 174699556 NBLA00301chr5 2805770 2806724 2808511 C5orf38 + 2805261 2808511 C5orf38 chr532747979 32750229 32823011 NPR3 + 32747421 32823011 NPR3 chr5 3787251637875378 37875539 GDNF − 37851509 37875539 GDNF chr5 63292326 63293075 −63292033 63293302 HTR1A chr5 76151227 76151876 76166895 F2RL1 + 7615058876166895 F2RL1 chr5 76968098 76969322 76970278 OTP − 76960292 76970278OTP chr5 140286488 140287347 140372113 PCDHA13 + 140286485 140372113PCDHAC1 chr5 140742560 140743119 140872730 PCDHGA6 + 140742650 140872730PCDHGA7 chr5 140747458 140748016 140872730 PCDHGA7 + 140747635 140872730PCDHGB4 chr5 140767256 140768329 140872730 PCDHGA9 + 140767953 140872730PCDHGB6 chr5 145698688 145700318 145700276 POU4F3 + 145698779 145700276POU4F3 chr5 157030866 157031611 + 157031138 157039738 C5orf52 chr5158690162 158691536 − 158674368 158690059 IL12B chr6 10994972 10995711 +10995049 11082527 SYCP2L chr6 26125798 26126758 − 26125238 26126019HIST1H1A chr6 27948718 27949880 27949268 HIST1H4L − 27948904 27949268HIST1H4L chr6 30050074 30050923 + 30050870 30054156 HCG9 chr6 3081875330820599 30820306 IER3 − 30818954 30820306 IER3 chr6 43720000 43721360 +43720787 43746722 RSPH9 chr6 78228956 78230415 − 78228666 78229839 HTR1Bchr6 83130068 83131208 + 83130257 83133335 TPBG chr6 112147783 112148610112301320 FYN − 112089177 112147958 FYN chr6 117192435 117193290117193579 FAM162B − 117180053 117193579 FAM162B chr6 137857127 137857980− 137855028 137857224 OLIG3 chr6 150326335 150328184 150336539 ULBP1 +150326835 150336539 ULBP1 chr7 526958 527907 − 503424 526007 PDGFA chr75077779 5078727 5079380 LOC389458 + 5078263 5079380 LOC389458 chr727100570 27102300 27102150 HOXA1 − 27099138 27102150 HOXA1 chr7 2715674927161918 27162821 HOXA7 − 27159862 27162821 HOXA7 chr7 27185469 27186510− 27176734 27186401 HOXA10 chr7 27190648 27192605 27195437 HOXA11AS +27191551 27195437 HOXA11AS chr7 27248657 27253218 27252717 EVX1 +27248688 27252717 EVX1 chr7 51352205 51352779 − 51051402 51352009 COBLchr7 116906858 116907422 + 116907252 117095954 CFTR chr7 120755525120756474 120768394 WNT16 + 120756325 120768394 WNT16 chr7 129912506129914030 + 129913281 129933367 MEST chr7 131983416 131984045 −131820060 131983987 PLXNA4 chr7 156125196 156126661 + 156126161156162581 RNF32 chr8 10626015 10628284 − 10618687 10625432 SOX7 chr823618422 23620876 23619867 NKX2-6 − 23615909 23619867 NKX2-6 chr824826911 24828465 24832511 NEFM + 24828359 24832511 NEFM chr8 2486888224870630 24870048 NEFL − 24864387 24870048 NEFL chr8 26777575 2678052126778839 ADRA1A − 26683138 26778839 ADRA1A chr8 57522214 57523567 −57516068 57521836 PENK chr8 65873955 65874718 − 65671082 65873902 CYP7B1chr8 71144440 71147178 71146116 PRDM14 − 71126576 71146116 PRDM14 chr898359200 98360130 − 98354889 98359352 TSPYL5 chr8 102573379 102574213 +102573843 102751128 GRHL2 chr8 110725080 110726354 110773196 GOLSYN −110655580 110726998 GOLSYN chr9 34369432 34371281 34387849 C9orf24 −34369016 34371598 C9orf24 chr9 85760867 85761816 − 85743046 85761483C9orf64 chr9 90339041 90340305 + 90339835 90380523 NXNL2 chrX 111211507111212571 111212660 TRPC5 + 111212908 111587129 ZCCHC16

TABLE 6 Genes with frequent hypomethylation at the 5′ end Annotated torefseq Peaks Average P value Average P value No. of Tumors with Gene chrStart End score in Normal score in Tumor hypomethylation Strand StartEnd Symbol chr1 55043525 55044789 9.92 0.90 29 + 55044323 55080525C1orf177 chr1 245760561 245761190 11.23 0.17 30 − 245754603 245760729LOC148824 chr10 105207770 105209419 8.15 0.76 29 − 105203133 105208638CALHM1 chr10 134590561 134591211 14.89 0.13 30 − 134471886 134590989C10orf92 chr10 135086692 135087332 6.87 0.00 30 − 135084159 135088111SPRN chr10 135228897 135229736 10.54 0.41 30 − 135218337 135229128 SYCE1chr11 50213739 50214494 20.02 0.70 30 − 50195574 50214199 LOC441601chr11 94439813 94440372 9.32 0.44 29 + 94439703 94444036 SFRS2B chr1250004030 50004674 6.53 0.00 28 − 49961088 50004205 BIN2 chr13 3568637835687238 8.02 0.00 30 − 35640346 35686752 SOHLH2 chr14 69107587 6910993615.81 0.35 30 − 69106284 69107671 C14orf162 chr15 22471456 2247240512.17 0.27 30 + 22471633 22479686 C15orf2 chr16 66571411 66572180 13.681.17 27 − 66567066 66571953 DPEP3 chr16 74126384 74127343 8.12 0.97 26 −74119928 74126569 CHST5 chr17 23658080 23658835 6.71 0.65 26 − 2362773423658144 FLJ40504 chr17 25927568 25928103 6.43 0.63 26 + 2592760825988611 LRRC37B2 chr18 27594502 27595268 19.93 0.67 30 − 2759365627594841 MCART2 chr18 59294602 59295373 7.41 0.45 27 + 59295123 59323298SERPINB5 chr19 16861645 16862379 14.99 0.95 30 + 16860825 16863830 F2RL3chr19 37902256 37902885 10.24 0.12 30 + 37902518 37973554 TDRD12 chr1951818645 51819269 8.01 0.00 30 − 51815564 51820194 PTGIR chr19 5365682053657569 21.31 0.64 30 + 53656317 53661179 KCNJ14 chr2 106450965106451409 14.15 0.78 30 − 106387567 106451233 RGPD3 chr2 107809732107810061 7.36 0.61 29 + 107809819 107875431 RGPD4 chr2 108702582108703121 13.69 0.59 30 + 108702368 108768699 RANBP2 chr2 131918756131919690 16.19 0.14 30 − 131916205 131918937 LOC401010 chr2 238371764238372800 28.42 0.80 30 + 238372126 238416189 RBM44 chr20 41765444177393 10.90 0.63 28 − 4149277 4177659 ADRA1D chr20 29689051 296897005.41 0.43 27 + 29689351 29696461 COX4I2 chr20 61963314 61964663 12.670.30 30 + 61963009 61964785 C20orf135 chr22 35231985 35232930 14.68 0.8030 − 35213178 35233094 FOXRED2 chr22 48869825 48871184 9.64 0.19 30 +48870620 48942243 MOV10L1 chr3 48673342 48674613 9.92 0.36 29 − 4864889948675352 CELSR3 chr5 177546540 177547089 11.17 0.15 30 − 177544116177547039 GMCL1L chr6 66554331 66554795 5.01 0.00 30 + 66554492 66556096MCART3P chr7 2684888 2685852 9.68 0.13 30 + 2685688 2721595 AMZ1 chr88597079 8598233 11.15 0.96 27 + 8597075 8599027 CLDN23 chr9 4194431041945459 13.06 1.11 26 − 41942398 41945076 MGC21881 chr9 4418546544186109 15.45 1.12 29 + 44185511 44246438 KGFLP1 chr9 66292609 6629387012.00 1.11 27 + 66293050 66295728 MGC21881 chrX 48050422 48051291 6.130.44 27 − 48039828 48050558 SSX9 chrX 48156111 48156755 8.29 0.68 29 −48146471 48156288 SSX4B chrX 52668418 52669072 5.11 0.26 25 + 5266870952679723 SSX8 chrX 52796355 52796804 8.08 0.48 29 + 52797032 52807342SSX2 chrX 55263129 55263764 10.17 0.23 30 + 55263516 55267266 PAGE5 chrX84075635 84076305 10.44 1.13 28 + 84075812 84076551 UBE2DNL

DNA SEQUENCE (from UC Santa Cruz GenomeBrowser, Human March 2006 (hg18) assembly) SEQ ID NO. 1:Chromosome 7, starting from 152252636 to 152253475CACACCTGCAAGCTGCGTGCAACCGAAAGGCTGGCGACCTGTAGCTTGTCAGATTCAGATATGGTGCAGGTGAGGGCCTGGCAAACATTTAGTTTAATTTCTGAGACCGTTAGAGACAAGGTTAAATTCCAGGTGAACGCGGTTCCAGAGTGGCTGGGATTCCCACTCTCAGATGGGAACCCTTCAGACAAGAGTAAAACAGGAGGGACTTCTTCGCCGCCCGCCCTGCGGTGTGTGCCACGGAGGCCGCTGGGTGGGGGCGTCCAAATGTGGAAGCGTTCCTCCAGCATGCGTGGATATCCTGAGGACTCACGGACACCGATTCCCTCTTCCGGGCCCAGACGTCCTAATTCCCGTCCCCCTTGAGGTGAGATCCCGTTTTCCCCCAGCCAAAGGGACTCCGGAGCCGCATTCACCTGACTTGGCTGCCAGCGCCCGCCTTCCTCGAGTTCTCAGGGCAAGGGGCTGTGGTGCATTTTCTGGGGATGGTTTGTGGATGTGGAAGAGGGAAATCCTTTCAGGGATTTTAGTCATTTGGCAATTGGTGATTTTCATATGACGTGCTGGTTTTAGGGGCCAACTCTGATAAGCTGGGACGCCCTCCCCTTGATTCTAGAACAGAAGGCATTTTGGGCTAAGGACGCTCACAACCCGGTTTCCTGTCCCGCGTGGTTCATGGGGCTTGCCCTGGGCCTGGGAGGCCGGTGGCGGGAGCCTCCCCTACAGTGAGGTGGGTGGGGGGCGCCACACGGCGTCTCAGGGAGAGGGGAGAGGCGGCTTCTGTTCCGGGAAGAGGGAACGGTGGAGGCCGCCGTTCTCTTGGGCGCGGCCTCTGCTGGG SEQ ID NO. 2:Chromosome 5, starting from 158456493 to 158457132CCAAGGCTCCCGGGCCTGCTCCCGAGCCGAGCCGGGGACGAGGGGCGACAGCGCTGCGCCCACTTACTTTTTCCTTCTCCACGAACCCCACAAACGCTGTCCTCTCGATCTCCACGGGCTGGCCCTGTCTGTCGTAGAGGGCCAGGACGAAGTGGAAGAAGTTGGATTTCCGCAGATTGGAAGGCGGCTGCTTCTCAAAGTGAGCCCGGGCCAGACCCACCCCGCTGCGGCCAAAGACGCAGAGTTAGATGGCTAAACCGGACGCCGACCCGCGCCCCTTGTCACCCTGTACACACACTCGAACCCTCAAACACCCACCTCCTCTTCCCCCAAAACATCCAGTGGGCGCTCTTCACGCCCCTTCAGGCGACATAGACCCAGCTGACAACGGGACACACATGTCCCCTCATGTCCTCGTCCTCTCCGCCAAGGACGGGTGCGTCCTCAGAACTCGCTGAAGAGGTGTGAAAGTTTTGTTTGGGTTTTGCGCGCGAGATGAAACTCTATAAGCATTTACTCCTACCTGACACGCACTGCGTGGGGAGGAAGAAAAGTGGCGGTGGATGAAGTGAACCCACCTTCGCGGAGTAGAACCCACAGTTATATTCCGGCACCCCTCGAATC TCACGTCTCAGCTAGCSEQ ID NO. 3: Chromosome 12, starting from 47771091 to 47771619CCTAGGGTGGCAACAGTACTACTGCAGACTCAGTTTCCCGGAGGGAGCCCCCTTTGGGCGGATTTTACCACCCTCCTCCTACTGTACCAGCTTTGACCGACACGTGGACGTGGTTGCGGGACTCGTAGTAGACCCAGTCGAAGCCGGCTTCCACTGCGAGGCGCGCCAGCAACCCATACTTGTTGCGGTCGCGGTCAGACGTAGTGATGTCCAAAGCACGGCCTTCGTAGTGGAGTGAATCCTGAGCGTGGTGGCCGTCCTCGTCCCAGCCCTCAGTCACTCGTAGGCGCACTCCGGGCCACATGTTCATCACGGCAATGGCCAAAGCGTTCACCCGCTCCTTACAACGCTGGCGGGGAATAAAGGAGTCAGTCTCCCCACCACCACCCTTGGGGCAAAAGGGACCTGGATAGGAGGGTTGATTCTTTGTTATTCCGGCCCTACTCTTACCCCTCCCAGCTTTTGAGTGTCCTGGAGAAATGAGAATCTGAGTCGATGGTAGTCACCAATCTGCACTCTGTTCCCAGG A SEQ ID NO. 4:Chromosome 14, starting from 22360909 to 22361365CTCAGGTGACCGGTCTGGGTGCGGCGTGGAGATGGCGAGAGCTAAGTGCTATGGCCGCGAAAGGGTGAAGGGCAGGGGAGGAAAAGGCCGAGGGGAGGCGAACGCTCAGGTTCACACATATGCAAGTGGGCTCTACAGCGGACTTCGAAGCATACACTCAACTCCCCACCATGCGCCGGCCCGCTGCTCTACCCCTGAAAAGCTCTCCCCTGGCCCGCGATCCTTGCTGGCCTACCCCTTGGTCGTTCCCATCCCCCTCTCCGCCCCCGCCCAACCTGCTGGCCTCACTCACCTTTGTCCCTTTCTCCCATCCCCACCCCCCCACCACCCTCGCTTTCTCCGGGCCCCTCCCTGTCGTTTGTCTCCCTTTTATTTCCCTCCTTCCTTTGGGCCCCCAGTCTGCGGCCCTAGGACAACGCCTTCCCAAGCTCGGACCCAGCCACCGTCCGCCTTCATT SEQ ID NO. 5:Chromosome 2, starting from 200041833 to 200043073CCCTGATCAGTTTGTATATGAAACACTTTTTTTCCTGCCTTAAGGTTGGAATCTGGTAAGTTTCAGACAAGTGGTTGCAGGGGGAGATGACCCAGGGGACCGTGCGGGACAGGGCCGGTGAACCCGATTCCGCGAGGCGGGCGTCTCGAAGGGACTGGTCTGCGTGCCTCACCGGGTTCCCCAGCCTGGCTCCGAGGAAAGGCCGCGCGGAGGCGAGAAAGGGCCCACATCTCGAGTCTCCAGTCTCTCCTTGAAAAGAGCACCCCCTAATCCCGGAGGAAGGGAGGGTGAAGGAGGAGGGGAGAGCCCACTACAGGCAAGGCAGGAAGGTTAATCATTTTGTTAACCGGAAAGGGTGCTGCCCCGCCTGGCCGTCGTCACCGAGCGTGTCAGTCACCCGCTCTCGATTTCGCCGCAGAATTACCTGCTCAAAGAGCCACCCCAAGAAGCCCTCAGCTCCGAGGTCTTCCCGCCCGTCGTCTGCGGGAAGTAAACGGCGTCCTCCCTCCAAAGAAGTGGATGCCCCTTCGACTCTCCTCCGCCCCCCGGCGGTTCCTAAATCTAGGAAAGCCAGCCCCGGCATCGCTCACTACTCCTGCACCTCAGCTCTGGGATACCCGCTCAGTACAGACAGACCATCAACCAAAAAAGAAAAGAAGAAAAGAAAAGTGGTGGTGGGGGTGGGGGTGGGCAGGGAGCGTAAAGTAGCGAGATCACTTATTTGACTCTTCTGTTTTCTTAGGAACCTGATCCTGGCCCCCGGCAGAGCATAAGGGCCCGAGGCCAGGGACCTAGAGGCCGGACCAAGTTCGCGCGAGGTGTTCACAGCCTAGGCGCCCCGGGCTTGGCTTCGTACTCTGCAACAAGCCAAGCTCCCTTGCCACGAGTATATCAGCAACTTCAGAAATAAACACCAGCGAAACCCCTTCCCACATTAGCTTGTTGCTGTGCTACTGACACTGGGCTTAGTCGATGTTCACACCTATTACATTTTTCCGGAAAGTGGGAGGAGCAGGGGGTAGAGAAAGCTGTGTGTATTGAGGGAAGGGGAGATGTGTGCAACACCTTCCAGAAAAATGGGGAGGTAGTTTAGAAAGTTTTTCTCTGTAACAGCACAGCTTCTGAGAAGGCAGAGAGAAGAAAGCAGACAGGAAACGAATCCCTTCCTCCTCTCCCCTTTACCCCCTTCCTTCACTCCCTAGACCAAGCTGGTCACCCGGCTGGGAATCAAGATTGTGTGG SEQ ID NO. 6:Chromosome 4, starting from 41443929 to 41444657AAAATGCCAAGAGCGTGCGCTCTAAGCCTCTCTCGAACGCACAGCCACACCAAATCCAGTATTTCTGATCGGCCATGGGGCCCTAGGTCCTTCTCACTCGAGGCTCCAGGACTTCGAATTTCACCAGCCGCCCCTCACCCCACACCTCCCCGGACCAGTGCGGCGGAGCGGGGTCGGTTTCCAGGCGCGCGTACCTGGACTCGCGCCTCTGTGAGGTCGATCTTCAGGGCCAGCTCCTCCCGAGTGTAGATGTCGGGGTAGTGAGTCTCCGCGAAGACCCTTTCCAGCTCTTTGAGCTGGGCACTGGTGAAAGTGGTGCGGATGCGCCGCTGCTTGCGCTTCTCGTTGAGGCCGCCGTGGTCCGTGAAGAGTTTGTAAGGAACTAGAGTATGACAGAGGAGACAGAAAGTGAGCAAATCAGCCGGCAGCTCGCCGGCCGTGGAGCTAGAATGTGAGGACTCCAAGGTCAGGTCATACGGCAGCCGCTACCTACACCCGCGCGAGAGAGTTGCCGAGAGAAGCTGCGGGAAGAAACCGAGGAAATTTGTTATCTGCGGAAACCTGGGCTCAAACTTCGGGCTTGGCGGAGTCCTTTCTGGCACAAGCGCCTTTGGGTGGATTGAAACAGCGCGATGTGACGGACTTGAGATAGTGCTTTCAGCAGGAAAAAACCAGAGAGGAAAAAAATCCAAAAACATCAGTCGGAATATCAGGGAGCCA GGAAAGAAG SEQ ID NO. 7:Chromosome 17, starting from 41329931 to 41330876ATAAATTTCACCCGTGTCCAAGTTCACCGCGCCCCCAAAACCGAGTCTGGGGCGGCAGGGGGAACTCCTGGCCAACGAATCCATGCCTCGCCCTCCTGTGATGAACCTGGTACGCACGGTTTTCTGGTTAATTCTATCGCTGAAAACTGGTGCGGGGGGCGCACTTCTGAGACGGAAGAGCATCTAGGAGCTGAATCCTCCACGCGGGTCGCCCAGGTTGATCTGAATTTCTGGGGAATGGCTTGGCTGCCCGCCCGGGACCAGGCCGACCCTCCTTGACGGTGGCGTAGAGGGCTGGAGCCTGGGTACTGCGAGGCTCCTCGCATGGCTGGGCCCGCCGCGAGGGGTTGCAGAGCGGCTCAGGGATCGATTCAAGCATCGTCTCTCCTCCCTCGCCCCCAGACAGAGCTGGGCGCGGGGTTCCCCTTCCAGATGGAGCGAGGGTCTCGGGGTGGCCCCGGAAAAGGGGAGCCCGCGGCCACGGCTACGTATTGCCATCTCGCGAGCAGAGATGTCACCTCCTGCCTTTGGAGGAAAGGGAGCCCGGTGGGGATGAGCGCATTTAGCCCAATGCTGGGAACAAAGCGCACTCCGCGCTTCTGCGATTTCGCTCCATTTTGAAATGTGTTGGCGCTTTGGTGGGGCCGCTGCGGTGGGCAAGGCCGGGGGCGCTGTTAATGGAGGAACCTCAGGGGGACGGTCCTTCGTAGGAAACTCTATCCTGGCTCTGCGCGCGCTTTAAGGAAATGGCTTCCCTCCAGGACCTCGAGGGATGCAGCTTTTGCGCGGATGACGGTGGGGTGCTGAACCAGCCGGTGCGCCTCTGGAAATGTCTGGGCACGGATCCTGGGGCCATCGACGACTCCTCCCCATTCCCAGCAGGCGGGAGCTCTTACATTCCGAGCGAGTGACCCCTCTCACCCTCTGGCGCTCACACACCTGTAAC SEQ ID NO. 8:Chromosome 19, starting from 56897971 to 56898600GTCAAGATGCCACCTGGCCGATGGACCCCGGGAAGCTGCATGGATCTCAAATGGGCCAAGGGCCCCGGGCTTGGAAGGGCAGGTGGCGCCTAGACCCAGCGTTCCCGCTCCACAGGTCTCCAGGAGTTCGCCCGGCGTTTCCTCTGCAGCGGGTGCTACTCTAGGGTCTGCGACCTCCCGCTGGACTGCCCAGGTGAGGGGGCGGGGCCTCGGGGTGCAGGAGGCCAACCTGAGAAAAGGGACCAGAGCACCGAGGGGCATAAGCTGGTGGCGAGGCCAGACGTGGTCGGGGCCCAGCGAGTGAACCCTGCTCCGTCTTCAGTTCAGGATGTGACAGTGACTCGGGGCGACCAGGCTATGTTTTCTTGCATCGTAAACTTCCAGCTGCCAAAGGAGGAGATCACCTATTCCTGGAAGTTCGCAGGAGGAGGTGTGAGTCGGGGCGGGGCCGGCGCGAAGAGTTTAGACGGGCGAGTTAGCCTTCACTAGAGGTTGGGGAGTCAGCTTGCGGGGACGGGACTCCGGGAATCTCCGTCTAGACACAAGCATCAGCAAGAGGAGGGTCGCGGGATTTAGGGGAGAGCTCGAAGATCAGAATGCAGGACCAAGGACCTGGGGCGATGG CTCACG SEQ ID NO. 9:Chromosome 20, starting from 44073717 to 44074646CTCCGACGGCTACCGCTGGTGCGCCACCACCGCCAACTACGACCGGGACAAGCTCTTCGGCTTCTGCCCGACCCGAGGTACCTCCACCCTGTCTACCAGGTTCAGCCCCGCCCTCTCATCATGTATTGGCCCCCAAAACGCGGCTCTTCCCTCCCATCAGTTTGTCTTTCCACTCTCATTGGTCCTCAGGACGACCGTGACTCCGCCCACCTACACCACATTTCCACCACTATCCCTGACTTCCAATGGCCCCGCCCCAGCCACTAAGGTTCGGCCTTTTCTGCCCAGCTGGCCGCCTCTTCCTTGGTCTGGTGTCCCAGGCACCGCCCACGGGTCTAGCCTCTTCTCAGGAGTGCTCTACAGCGCCCCCTAGGCCACCAAGATTGTTTAGCTCCCTGTCGGGTCGGCCCCTGACTCCTTATTGGACTCATCCATCTGGCTCATCCAAGGCCTTGGGTCTCTCCAGCTGACTCGACGGTGATGGGGGGCAACTCGGCGGGGGAGCTGTGCGTCTTCCCCTTCACTTTCCTGGGTAAGGAGTACTCGACCTGTACCAGCGAGGGCCGCGGAGATGGGCGCCTCTGGTGCGCTACCACCTCGAACTTTGACAGCGACAAGAAGTGGGGCTTCTGCCCGGACCAAGGTAGGCGTGGTCCCGCGGCTCCGGGGCTGGGGTTCCCGGCAGTGGTGGTGGTGGGGTGGCCAGGGCTGGGGGCTCGGCCCGGCGCTCACGTCTCAGGCTCCCTCTCCCTCCAGGATACAGTTTGTTCCTCGTGGCGGCGCATGAGTTCGGCCACGCGCTGGGCTTAGATCATTCCTCAGTGCCGGAGGCGCTCATGTACCCTATGTACCGCTTCACTGAGGGGCCCCCCTTGCATAAGGACGACGTGAATGGCATCCGGCACCTCTATG GTGAGGCAGGGGCAGGGASEQ ID NO. 10: Chromosome 8, starting from 26777575 to 26780521CGATGCCCAGCGTTTTGGCCGCTTTCTTCTCCCGGGAGAACTTGAGGAGCCTCACTGAGAAGTGCGTCTTGGTCTTGGCGCTGGCCATCCCGCTGCCTCCTGCCGGGGCGTTTTTCCGATGGATGCGGAGCGTCACTTGCTCCGAGTCCGACTTGTCGGTCTTGAGGCCAGACTTGAGGCCCCGGCTCTCCCTCTTGGCCACCACGTAGACGCGGCAGTACATGACCAGGATGATGGCCAGAGGCAGGTAGAAGGAGCCCAGCGCTGAGAAGAGCACGTAGCCCGGCTCCTCGTTGATCTGGCAGATGGTCTCGTCCTCGGGGGCCGGCTGCCTCCAGCCGAACAGGGGTCCAATGGATATGACCAGGGAGAGTGCCCAGACGCAGAGCAGAGCCATGAGACCCCTCCTCTGGGTGACGATGGTTGGGTAGCGCAGCGGGTAGCTCACGCCGATGTAGCGGTCGATGGAGATGATGCAGAGGCCCATGATGGACGCGGTGCAGCACAGCACATCCACTGCCGCCCAGATGTTGCAGAAGACCCTGCCGAAGGCCCAGTAGCCTAGGACCTCGAAGATGGCGGAGAAGGGCAGCACCGTGGAGGTGAGCAGGAGGTCGGCCACCGCCAGGTTGACGATGTAGTAGTGCGTGACTGAGTGCAGGTGTCGGTGACAGGCTACGGAGAGGATCACTAGGATGTTACCCAGCACCCCGAAAAGAATGAGGCCCCCCAAGATCACCCCGAGCAGAATGGCCTTGGAAATGTTCACCGGTGCCGGCGGTTGGGTGCAGTTGGAGCTGTCGGAAGCATTTCCCGAGAGAAACACCATGGTCCCAGCCGGGGCCGGGCGAGGTCCGGCTGTCCAGGGCCACCTCCCGGGCTGGCGCGGAGGCGGGAGCGCGGGAGCCGGGAATCAAAAGGTCTCGGCTGGAGGGAGCCCTGCCAGGTGGGTTTGGCTGGGGGTGAGAGCGCGCGCGCGGGTGGGAAACAACCCTGGCCAGCCCTGGGAACCCTCAGAAGGCCACATGAAGGGGCAGGGCATTAAAGACATGGCCAGGGGCGCGCGGGGCTGCCGGGGACCCTCTCCACCTGCCGGGCTGGCCTAGCCCGGGACCCGGACTCCCTCCCTACCCGAAGCTGGGTGCGAAGATCCAGGAGACTCCTTGCAACATGCAATTCCAGAATTACGAGAATCTGCTTTTCCGCGCTGTCTTATTCGTCCTGGCTGGGGGAAGATTCAGCATTCTGCACATGATTCGGAATTCAAAACTCCAGCGCCAGTCTCTCCCTCAAACCAAAAGATCAGCCGTCGACGCTCAAAGGCAGGGACCGATGGTTGGGTAGCGCAGCGCTACAGGTTGGGCGCTGCTCCTGGCCCGCAGTTACCTACATTTTGAGCTGCCCCACCGAAGGCTCCTGCTCTCTCCAGCTTCTAGGAGCACAGGTCAGGGGACGTAGGTGTGGAATATGTGCTGAGACCCAGGAGGCTGCGGGGCATCGTTTGACCGCGTCCACCTGAAAGAGCGCAAAGAGAAAGGCGGCTTTGAGCTAGGCGCCCCAGGGAAAGAGGCTGTGCTGAGCTTGACGGGTTGGGGGACACCAGTTGGGAGCCGGGTTGGTTCTGCGGTCCAGAAGCTGCTTCGCCCGGCAGCGGTGGAGGCGACTTCGGAGCTCATCTCGCGCCCCCACCACTGGGAACCTGCCTAGCGCACTCTACTGAGTCACCTCTGCCCGAGTTCAGGATCCGAAGCGAAAAACAAACAAAAAAACAAATCCCCAAAACCCCAGGCTCCAGCGCTCGAAGTCTGGATTTCGAGCGCAGGTACCACGAAATTAAAATCCTCGAAGCCCCGGAGGGGCGACTGCGGCTGGCGAGTGTGTCGCACGGATCCTAGCCGGGGAATTCTGGAGGATGTACTCGGTTTCCGTTAGAACAGGTTGCCTCGCAGTCACCTGGAGGGGGCGGCCTGGGAGAGGGAACAGCTGCGACCCAGGGACCTCAGGGCCAGGACCACGAGCACGCTCACTCTCACGCCGGTGGAATTCGCACTCGGGTGTGCAGAGCGCACCGGTCTGTCCACCAGCCAAGCTGGCTTGAGAGCCAGGTCCCGAGCGAAGCCGGGAGGGACCCGAAGACAGAAAGCGACCCAGGTCTGTCCACGACGCCTTTCCAAGCCTCACTGTTGGCCACCCGAGGGGCTCCAGCTGCCAGCCGTAGCGGGACTGGCGGGGGACGGGGTGGGGAGGACGTCCCTCTGCAGAGGGAGCTTTGCAAGTGACAGTCACTGCGGATTTTGCAGGCGTAAACATTAAGCCGGCATGCCAGCGGGCAGGGGCCGCCTTCGATTTTCTGGGCTGGGGGCCACCATCTGCGTAAGAAACCTGGGTTTCCACAACTGCGAACCTACCGCAGGGACTCGGCAGGACAGCGCGGCGTGAGGAAGTGCCCACCTTGCCTTCTGGGGTTGGGCCCTGAGGTGGGACCTCGGGCAAAGACTCTTGTTAAAATCGCAAGTTGGAGACCTCACAGGTGGTATTAAAAACGTGCCACTGCAGAAACCCTTTTCCTCCTACCTCGAGGGTGATAAACACAGGGTTAAAAAGACACTTGTTCAGTAGAAGGCAACTTCGCAGAAGATGTAAGGGAATCGGGGGTGGGGTAGAGGGGCCGGTATAAAACCTGGTGGAAAAAGCGGGAGGCGCTGGGAAAAGTGGGGGTTCCGTCTCACCAGACGGCGGGGGAGGTCTGCCCTCACCCACTCGGCCCTGCGGGACGCCGGCCCCGGCGCACTCACCTGAAGCGCCGCTGCTGAGCCACCAGCTCGCGCGCGGGGGATGTGGACCCGGCTTCGGTCCCGGGAGCTGCCTGCCTGCTCTTCTCTGGAGGCGGAGAGGGGACCGTGTCTCCGAGGCACCAAATCCTTGTCC TTTTGCACCCGCTGACTCA

1. A composition of matter comprising a DNA hypermethylation braincancer marker on acellular DNA of a human subject selected from thegroup consisting of SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO.4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8, SEQ ID NO. 9,SEQ ID NO. 10, and combinations thereof.
 2. A method of diagnosing abrain tumor in a human subject comprising: providing a sample containingacellular DNA from the human subject; determining whether one or moreDNA hypermethylation markers are methylated on the acellular DNA; anddetecting the methylation of the DNA hypermethylation markers isindicative of brain cancer.
 3. The method of claim 2, wherein the DNAhypermethylation markers are selected from the group consisting of SEQID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ IDNO. 6, SEQ ID NO. 7, SEQ ID NO. 8, SEQ ID NO. 9, SEQ ID NO. 10, andcombinations thereof.
 4. The method of claim 2, wherein the DNAhypermethylation marker comprises CpG clusters associated with homeoboxgenes.
 5. The method of claim 4, wherein the homeobox genes are selectedfrom the group consisting of HOX genes clusters HOXA, HOXB, HOXC, HOXD,the homeobox genes DLX1, BARHL2, PITX2, developmental transcriptionfactor gene simple-minded one (SIM1), NKX2-8 PAX9 and FOXA1.
 6. Themethod of claim 2, wherein the brain cancer diagnosed is a brain tumor.7. The method of claim 6, wherein the brain tumor is a grade I tumor. 8.The method of claim 2, wherein the sample is selected from the groupconsisting of a blood sample, serum sample and plasma sample.
 9. Themethod of claim 3, wherein the brain cancer diagnosed is a brain tumor.10. The method of claim 9, wherein the brain tumor is a grade I tumor.11. The method of claim 3, wherein the sample is selected from the groupconsisting of a blood sample, serum sample and plasma sample.
 12. Themethod of claim 4, wherein the brain cancer diagnosed is a brain tumor.13. The method of claim 12, wherein the brain tumor is a grade I tumor.14. The method of claim 4, wherein the sample is selected from the groupconsisting of a blood sample, serum sample and plasma sample.
 15. Themethod of claim 5, wherein the brain cancer diagnosed is a brain tumor.16. The method of claim 15, wherein the brain tumor is a grade I tumor.17. The method of claim 5, wherein the sample is selected from the groupconsisting of a blood sample, serum sample and plasma sample.
 18. Themethod of claim 6, wherein the sample is selected from the groupconsisting of a blood sample, serum sample and plasma sample.
 19. Themethod of claim 9, wherein the sample is selected from the groupconsisting of a blood sample, serum sample and plasma sample.
 20. Themethod of claim 12, wherein the sample is selected from the groupconsisting of a blood sample, serum sample and plasma sample.